Integrated Solutions
for Nucleic
Acid
Purification
Table of Contents
Genomic DNA
3
Plasmid DNA
15
Fragment DNA
25
Total RNA
35
You have a
choice for Nucleic
Acid Purification
• The yield and
concentration you need
• Application-tested protocols
• Mini → Mega scale
• Manual or Automated format
• Global support and service
DNA and RNA Purification Systems
from scientists for scientists, for all
your perfomance and throughput needs.
2
...from a broad range of sample types with
easy-to-use, labor saving systems.
Genomic DNA
Purify High-Quality DNA...
Sample types:
• Blood
Promega genomic DNA purification systems provide:
• Increased productivity: Save time and labor with our easy to use protocols.
• Yield: Our systems and methods provide high DNA yields and purity.
• Performance: Sample-type specific protocol proven for PCR/genotyping.
• Choice of throughput: Select the format (manual single-tube, manual plate
or automated plate/tube) to meet your requirements.
• Bacteria
• Mouse tail and animal tissue
• Tissue culture cells
• Plants
• Food
• Applications for other sample types
• Convenience: Choose a kit that processes multiple sample types or for
challenging samples, a specific sample-type kit.
How can we help you achieve your goals today?
A Comparison of Genomic DNA Purification Systems.
Blood
Mouse Tails Tissue
and Animal Culture
Bacteria
Tissue
Cells
Plant
Tissue or Manual Automated
Spin
Food
Handling Handling Protocol
Vacuum Magnetic
Protocol Technology
Wizard® Genomic DNA Purification System (Cat.# A1120, A1125, A1620)
Wizard® SV Genomic DNA Purification System (Cat.# A2360, A2361)
Wizard® SV 96 Genomic DNA Purification System (Cat.# A2370, A2371)
MagneSil® Blood Genomic, Max Yield System (Cat.# MD1360)
MagneSil® ONE, Fixed Yield Blood Genomic System (Cat.# MD1370)
Wizard® Magnetic 96 DNA Plant System (Cat.# FF3760, FF3761)
MagneSil® Genomic, Fixed Tissue System (Cat.# MD1490)
MagneSil® KF, Genomic System (Cat.# MD1460)
Wizard® Magnetic DNA Purification System for Food (Cat.# FF3750, FF3751)
3
Wizard® Genomic DNA Purification Kit
High molecular weight gDNA from multiple sample
types in a scalable format.
Product
Wizard® Genomic
DNA Purification Kit
The Wizard Genomic DNA Purification Kit provides:
• Confidence: Proven “salting out” method - long track record.
For Laboratory Use.
• Scalability: Convenient for any sample size.
• Value: Cost per prep based on your sample size.
M
Cat.#
100 isolations x 300µl
500 isolations x 300µl
100 isolations x 10ml
1
+
300µl
100µl
2
3
5
4
6
50µl
7
8
A1120
A1125
A1620
20µl
9
bp
10,000 –
3,000 –
1,000 –
• Safe and quick: Non-organic solvent method can be completed in
about 1 hour.
DNA Yields from Various Starting Materials.
Source
Whole Blood
Tissue Culture Cells
Animal Tissue
Mouse Liver
Mouse Tail
Insect Cells
Amount of
Starting Material
300µl
3ml
10ml
96-well plate, 50µl/well
Typical
DNA Yield
5–15µg
25–50µg
250–500µg
0.2–0.7µg
106–107 cells
5–30µg
11mg
0.5–1cm of tail
15–20µg
10–30µg
PCR analysis of genomic DNA isolated from decreasing amounts of whole blood.
PCR products from 5µl of genomic DNA isolated using the Wizard Genomic DNA Purification Kit
as the starting template. Lane M, 1kb DNA Ladder (Cat.# G5711); lane 1, no DNA control (–);
lane 2, Human Genomic DNA (Cat.# G3041) positive control (+). Lanes 3, 5, 7 and 9 contain
samples that were rehydrated at room temperature for 20 minutes. Lanes 4, 6, 8 and 10 contain
samples rehydrated at 4°C overnight.
Sample
Add Cell Lysis
Solution. Incubate.
5 × 106 cells
16µg
Plant Leaf Tissue
40mg
7–12µg
Centrifuge.
Bacterial Culture*
108–1010 cells
5–20µg
Yeast*
1.9 × 108 cells
4.5–6.5µg
Discard supernatant.
Vortex pellet. Add
Nuclei Lysis Solution.
Mix. Add Protein
Precipitation Solution.
*Overnight culture.
Centrifuge.
Transfer supernatant
to new tube containing
isopropanol.
Centrifuge.
Discard supernatant.
Add ethanol.
Centrifuge.
Aspirate ethanol.
Air-dry pellet.
Rehydrate DNA.
Flow chart of genomic DNA purification using the Wizard Genomic DNA
Purification Kit.
4
10
2862TB02_0A
• Performance: High-molecular weight DNA suitable for
amplification, restriction enzyme digestion, and southern blotting
from a wide range of starting materials.
Size
Easy-to-use gDNA isolation from cells or tissue in
20 minutes.
Product
Wizard® SV Genomic
DNA Purification System
The Wizard SV Genomic DNA Purification System provides:
• Speed: PCR-ready genomic DNA in as little as 20 minutes after
lysis.
Genomic DNA
Purification from
Mouse Tail Clipping
or Tissue Sample
• Flexibility: Purify genomic DNA from 20mg of tissue, 1.2cm
mouse tail or 5 x 106 cultured cells in a single prep.
Mouse tail clipping
or tissue sample
Proteinase K
(Cat.# V3021)*
digestion in
Digestion Solution.
• Yield: Purify 20–30µg of gDNA per prep from 1.2cm mouse tail
clips with an A260/A280 ratio of 1.7.
M
–
+
Cat.#
50 preps
250 preps
A2360
A2361
Genomic DNA
Purification from
Tissue Culture Cells
Wash tissue
culture cells
with 1X PBS.
Incubate at 55°C
overnight
(16–18 hours).
Vacuum
Ta
il
Li
ve
r
H
ea
rt
Br
ai
n
Ta
il
Li
ve
r
H
ea
rt
Br
ai
n
Spin
Size
Genomic DNA
Wizard® SV Genomic DNA Purification System
Add Wizard®
SV Lysis Buffer.
Add Wizard®
SV Lysis Buffer.
Transfer lystate to minicolumn.
3710TB04_2A
Centrifuge.
Amplification of genomic DNA isolated from various tissue sources using the Wizard SV
Genomic DNA Purification System. One microliter of purified genomic DNA from the indicated
tissues was amplified using PCR Master Mix(a) (Cat.# M7502) and mouse-specific IL-1ß primers
(1.2kb product). Reactions with Mouse Genomic DNA (Cat.# G3091; +) and without DNA (–) were
performed as positive and negative controls, respectively.
Vacuum Adapter
(Cat.# A1331)*
Vac-Man®
Laboratory
Vacuum Manifold
(Cat.# A7231)*
Bind DNA.
Wash, removing
solution by
centrifugation
or vacuum.
Transfer spin
column to a 1.5ml
microcentrifuge
tube (not provided).
Centrifuge.
3643MA02_2A
Elute genomic DNA.
Flowchart of genomic DNA purification using the Wizard SV Genomic DNA
Purification System. *May be purchased separately.
(a)
The PCR process is covered by patents issued and applicable in certain countries*. Promega does not encourage or
support the unauthorized or unlicensed use of the PCR process.
*In the U.S., effective March 29, 2005, U.S. Pat. Nos. 4,683,195, 4,965,188 and 4,683,202 will expire. In Europe, effective
March 28, 2006, European Pat. Nos. 201,184 and 200,362 will expire.
5
Wizard® SV 96 Genomic DNA Purification System
Easy-to-use rapid 96-well isolation of gDNA from cells
or tissue.
Product
Wizard® SV 96 Genomic
DNA Purification System
Size
Cat.#
1 x 96 preps
4 x 96 preps
A2370
A2371
The Wizard SV 96 Genomic DNA Purification System provides:
• Performance: Purified DNA PCR ready.
• Time savings: 96-well processing using either manual or
automated purification protocols for Beckman Coulter Biomek®
instruments
• Flexibility: Purify genomic DNA from up to 20mg of tissue, 1.2cm
of mouse tail or 5 × 106 cultured cells per well.
• Convenient: Purified genomic DNA from 96 mouse tails in 45–60
minutes, and from cultured cells in 30 minutes after lysis. No
centrifugation step required. PCR-ready gDNA can be isolated
from tissue or mouse tails directly from lysate.
Average Yield of Genomic DNA Purified from Various Sources Using
the Wizard SV and SV 96 Genomic DNA Purification Systems.
Amount of
Starting Material
20mg
Average
DNA Yield
20µg
Mouse Liver
20mg
15µg
Mouse Heart
20mg
10µg
Mouse Brain
20mg
6µg
CHO Cells
1 × 106 cells
5µg
NIH3T3 Cells
1 × 106 cells
9µg
106
8µg
Source
Mouse Tail Clipping
293 Cells
1
2
3
4
5
6
7
8
9
1
1
1×
cells
1
Add sample
lysate.
A
B
C
D
Bind DNA.
E
F
G
H
Mouse Tail Clippings
Water Only
M –C +C C4 C5 C6 C7 C8 D4 D5 D6 D7 D8
M
E4 E5 E6 E7 E8
3712TA04_2A
Wash.
No cross-contamination. Genomic DNA purified from mouse tail clipping samples or water samples were
purified in adjacent wells of a 96-well plate. PCR products were amplified from 1µl of purified sample from each
well using primers for mouse IL-1β. Ten microliters of PCR product were run on a 1.5% agarose gel and
visualized by staining with ethidium bromide. Expected PCR product from mouse tail clippings for IL-1β is
approximately 1.2kb. No PCR product was detected from water samples.
Elute genomic
DNA.
3637MA02_2A
Highly pure
genomic DNA.
Flowchart of 96-well genomic DNA purification using the Wizard SV 96 Genomic
DNA Purification System.
6
MagneSil® Blood Genomic, Max Yield System
Product
MagneSil® Blood Genomic,
Max Yield System
Size
Cat.#
1 x 96 preps
Genomic DNA
Walk-away, high-throughput, automated gDNA
isolation from small volumes of blood in 1.5 hours!
MD1360
For Laboratory Use.
The MagneSil Blood Genomic, Max Yield System provides:
• Efficiency: Save labor using our walk-away high-throughput
96-well purification method.
M
+
–
1
2
3
4
5
6
7
8
9
bp
• Consistent results: Maximum yield of 4–6µg of purified highquality genomic DNA from well-to-well and plate-to-plate.
– 318
• Time savings: Standardized protocol provides reproducible results
in 1.5 hours. Let us program your robot – downloadable methods
available at: www.promega.com/automethods
XX
P1
3rd Tips
XX
Blood Sample
Resin Disp
Heat Block
XX
XX
XX
Heat Block
P10
Orbital1
Lysis Wash
Lysis Disp
Elution Solution
PCR analysis of genomic DNA isolated from 200µl whole blood samples. PCR
products from 5µl of genomic DNA isolated using the MagneSil Blood Genomic, Max
Yield System. Lane M, PCR markers (Cat.# G3161); (+) positive control apc; (–) no
DNA control. Lanes 1–9 contain PCR amplification product from purified human
genomic DNA.
P7
Lysis Resin
MagnaBot®
XX
XX
XX
XX
XX
P4
P7
P11
Wash Plate
Salt Wash
P14
Salt Disp
P18
Elution Disp
SWAP
XX
XX
XX
XX
P2
P5
P8
P12
P15
P19
4th Tips
Elution Tips
Alcohol Wash
Alcohol Disp
XX
XX
XX
XX
P16
P20
P3
P6
P9
P13
Deck layout for the MagneSil Blood Genomic Max Yield System on the Beckman Coulter
Biomek® FX.
M 1
4°C
2 3
4
21°C
5 6
7
–20°C
8 9
4235TA07_3A
2nd Tips
WS1
4152MA05_3A
Cleanup Tips
Purification of apoptotic DNA using the MagneSil Blood Genomic, Max Yield
System. In some cases, blood samples that are stored or shipped at elevated
temperatures contain white blood cells undergoing apoptosis. The MagneSil Blood
Genomic System purifies DNA across the broad size range seen in a typical apoptotic
ladder. Genomic DNA was isolated from blood that had been stored at the indicated
temperatures for 8 days.
7
MagneSil® ONE, Fixed Yield Blood Genomic System
Walk-away, high-throughput fixed-yield genomic DNA
isolation from small volumes of blood in 1 hour!
Product
MagneSil® ONE,
Fixed Yield Blood Genomic System
Size
Cat.#
1 × 96 preps
MD1370
For Laboratory Use.
17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36
• Time savings: Complete walk-away high-throughput 96-well takes
1-hour.
37 38 39 40 41 42 43 44 45 46 47 48
Cleanup Tips
Heat Block
SWAP
P5
P9
P13
HeatXX
Block
P1
Wash Tips
Wash Solution
Alcohol Disp
Alcohol Disp
WS1
XX
XX
XX
XX
XX
P2
Working Plate
P6
Lysis Wash
P10
Lysis Disp
P14
Elution Tips
XX
XX
XX
XX
P3
P7
P11
P15
Resin Disp
MagnaBot®
Lysis Resin
Elution Solution
Elution Disp
XX
XX
XX
XX
XX
Orbital1
P4
P8
P12
P16
TR1
Magbead
Shaker1
4153MA05_3A
SC1
P17
Deck layout for the MagneSil ONE Fixed Yield Blood Genomic System on the Beckman Coulter
Biomek® FX.
4231TA07_3A
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
• Consistent results: Obtain 1µg (fixed yield) of highly pure DNA
from 60µl of anti-coagulated blood.
Place the blood samples onto the instrument and come back in an
hour to purified and normalized DNA ready for PCR!
8
ng control
25
0
12
5
63
31
The MagneSil ONE, Fixed Yield Blood Genomic System provides:
• Labor savings: Normalized fixed yield of 1µg eliminates the need
for DNA quantitation, reducing the number of steps needed for
genotyping.
Consistent yield of genomic DNA purified with the MagneSil ONE, Fixed Yield
System. Genomic DNA standards were loaded onto the gel at the following
concentrations: 250, 125, 63 and 31ng. Forty-eight blood samples were purified with
the MagneSil ONE System, and 10µl of the 200µl total elution volume for each sample
was loaded on the gel and visualized by ethidium bromide staining.
Easy, labor-saving 96-well isolation of high-quality
plant gDNA for PCR-based genotyping
Wizard Magnetic 96 DNA Plant System provides:
• Time and labor savings: High-throughput manual or automated
96-well protocols give freedom from the time and labor-intensive
steps normally associated with plant genomic DNA preps. Manual
protocol takes about 45 minutes while the Beckman Coulter
Biomek® FX robot will complete purification in about 25 minutes.
Product
Wizard® Magnetic 96 DNA
Plant System
Size
Cat.#
2 x 96 preps
4 x 96 preps
FF3760
FF3761
Place fresh leaf or seed sample
in 96 deep-well plate in the presence
of Lysis Buffer A. Add 1 or 2
grinding beads.
• Consistent DNA quality: From well-to-well, plate-to-plate across a
broad variety of plant material.
Grind. Centrifuge.
• Ease of use: Fewer steps and non-clogging magnetic particle
technology.
Transfer supernatant to a clean
plate. Add Lysis Buffer B/MagneSil®
Paramagnetic Particle mixture and mix well.
Incubate.
• Adaptable chemistry: Scalable purification for standard or deep
well plates. Modified chemistry for fixed yield purification and
direct genotyping of gDNA.
Genomic DNA
Wizard® Magnetic 96 DNA Plant System
Place the plate on the MagnaBot®
96 Magnetic Separation Device.
Discard liquid.
Time Required to Isolate gDNA from Plant Tissue Using Three Listed Manual
Procedures.
Wizard Magnetic 96 DNA Plant System
Commercial membrane plate
CTAB homebrew method
Steps
11
17
16
Time
40 minutes
55 minutes
135 minutes
Examples of Plant Sample Types Successfully Processed Using the Wizard
Magnetic 96 DNA Plant System.
Arabidopsis
Cotton seed
Soybean
Cabbage seed
Grass seed
Squash
Canola leaf
Green pepper seed
Squash seed
Canola seed
Lettuce
Strawberry leaf*
Carrot seed
Milkweed leaf
Sunflower seed
Chicory leaf
Potato tuber
Tobacco seedling
Chives
Radish leaf
Tomato leaf
Corn leaf
Rice leaf
Tomato seed
Cotton leaf*
Sorghum
Watermelon seed
* These samples need the addition of PVPP to the lysis buffer to remove phenolic compounds that inhibit PCR.
Add Wash Buffer and mix.
Place the plate on the MagnaBot®
96 Magnetic Separation Device.
Discard liquid. Repeat the wash.
Dry the particles.
Add Nuclease-Free Water.
Place the plate on the MagnaBot®
96 Magnetic Separation Device.
Remove purified DNA to a fresh plate.
3220M A 01_1A
Method
Flowchart of plant genomic DNA purification using the Wizard Magnetic 96 DNA
Plant System.
9
MagneSil® Genomic, Fixed Tissue System
Easy-to-use isolation of high-quality DNA from fixedtissue and paraffin-embedded samples
Product
MagneSil® Genomic,
Fixed Tissue System
Size
Cat.#
100 samples
MD1490
For Laboratory Use.
The MagneSil Genomic, Fixed Tissue System provides:
• Ease of use: No deparaffinization step with organic solvents. A
manual method to purify DNA from thin sections of formalin-fixed,
paraffin-embedded samples.
Add Incubation Buffer/ Proteinase K
solution to tissue sample.
Incubate overnight at 56°C.
• High-quality DNA: Demonstrated amplification of fragments as
large as 1,800bp.
Add Lysis Buffer and Resin. Vortex.
• Quick results: Proteinase K treated sample processed to purified
DNA in under 30 minutes, 6–12 samples in under 1 hour.
• Consistent amplification: Ultraclean DNA free of inhibitors of
amplification including very small fragments of DNA.
Place on Magnetic Stand
and discard solution.
A.
Wash with Lysis Buffer.
Wash with prepared 1X Wash
Buffer. Repeat twice for a total
of three washes.
B.
926 base
Size
Marker
972 base
Amelogenin
Fragment
Air dry Resin.
100 200 300 400 500
600
APC
1.8Kb
Fragment
Analysis of DNA purified from paraffin-embedded, formalin-fixed 10µm thin sections using the
MagneSil Genomic, Fixed Tissue System. Purified DNA was amplified and then analyzed on an
ABI PRISM® 3100 Genetic Analyzer. Panel A. Amplification with a set of 16 fluorescently labeled
primers. Amplification products range in size from 104 to 420 bases. Panel B. A 972-base
fragment amplified using an amelogenin primer set. Panel C. A 1.8kb fragment amplified from
the APC gene. Results will vary depending on the degree of cross-linking due to formalin fixation.
65°C
Add Elution Buffer. Heat.
Remove tubes from heat, vortex,
and place in Magnetic Stand.
Transfer DNA to container of choice.
3742MA06_2A
Size Markers
3931TB12_2A
C.
Flowchart of DNA isolation from formalin-fixed paraffin embedded tissue using the
MagneSil Genomic, Fixed Tissue System.
10
Walk away, low- to moderate-throughput, automated
genomic DNA isolation in 25 minutes
Product
MagneSil® KF,
Genomic System
Size
Cat.#
200 preps
MD1460
Genomic DNA
MagneSil® KF, Genomic System
For Laboratory Use.
The MagneSil KF, Genomic System provides:
• Labor savings: Walk-away automation of 1–15 sample batch
genomic DNA purification frees-up time for important work and
cuts labor costs.
1 2
5
6 7
8 9 10 11 12 13 14 15 16 17
4212TA06_3A
• Consistent results: Well-to-well 2–6µg of high-quality DNA per
prep from 200µl of anti-coagulated whole blood.
3 4
• Simple method: One instrument and one chemistry for gDNA
purification, no training or modification of protocols necessary.
Yield gel of genomic DNA isolated from 200µl of liquid blood. Fifteen representative
samples (single instrument run) of DNA purified using the MagneSil KF Genomic
System from blood. Ten microliters of the 200µl total elution volume were loaded on
the gel and visualized by ethidium bromide staining. Lanes 6 and 12 left intentionally
blank.
4213TA06_3A
• Save time: Automated purification method takes 25 minutes to
complete.
Multiplex amplification of Factor II (345bp) and Factor V (225bp). Genomic DNA purified from
200µl human whole blood as recommended in Promega Technical Bulletin #TB322 MagneSil KF,
Genomic System. Figure courtesy Dr. Greg Tsongalis, Hartford Hospital, USA.
11
Wizard® Magnetic DNA Purification System for Food
Labor-saving isolation of high-quality gDNA
from Food
Product
Wizard® Magnetic DNA
Purification System for Food
Wizard Magnetic DNA Purification System for Food provides:
• Ease of use: Protocol eliminates organic extraction.
Size
Cat.#
200 preps
400 preps
FF3750
FF3751
Food Sample
• Time savings: No long incubations or centrifugation steps.
• Reliability: Quality controlled for PCR-based testing for GMO.
Prepare Lysate.
• Flexibility: Scaleable manual method to purify DNA from 200mg
to 5g food samples.
Centrifuge.
• Consistent results: Ultraclean gDNA of a broad size range free of
inhibitors of amplification.
Bind DNA.
GMO Maize (%)
M
bp
0.0
0.1
0.5
2.0
10,000 –
Capture MagneSil® PMPs.
1,000 –
3019TA06_3A
500 –
300 –
150 –
1
2
3
4
5
6
7
8
9
10
Wash MagneSil® PMPs.
Dry particles.
Duplicate Fluka maize reference standard maize samples containing up to 2% GMO maize
were processed with the Wizard Magnetic DNA Purification System for Food. Ten percent of
100µl eluates of DNA in duplicate were loaded on a 1% agarose gel. The 0% standard shows a
greater yield and higher molecular weight DNA because non-GMO maize undergoes minimal
processing in the preparation of the reference standard. Lanes 1 and 2, 1kb DNA Step Ladder
(Cat.# G6941) and PCR Markers (Cat.# G3161), respectively.
S
S
C
O
Capture particles.
–
N
35
–
–
S
O
S
C
N
–
–
C
35
–
N
O
S
S
Corn Seed
(GMO–)
–
–
–
–
C
35
O
N
–
M
–
bp
35
S
S
Cornmeal
(GMO+)
Add sterile water.
Vortex and incubate.
1,000 –
500 –
300 –
195bp (S35)
180bp (NOS)
M
1
2
3
4
5
6
7
8
9
10 11 12
DNA isolated from cornmeal and ground corn seed using the Wizard Magnetic DNA
Purification System for Food. Detection of GMO sequences by PCR was performed using 1µl of
DNA sample per template. The first three replicate sample sets (lanes 1–9) are from GMO+
cornmeal. Samples represented in lanes 10–12 are from GMO– corn seed; nonspecific bands are
seen in these 35S and NOS lanes. Lane M, PCR Markers (Cat.# G3161). Lanes labeled C:
chloroplast PCR positive controls.
12
Elute DNA.
3013MA07_0A
3016TA06_3A
150 –
Flowchart of genomic DNA purification from food using the Wizard Magnetic DNA
Purification System for Food.
Product
Wizard® Genomic
DNA Purification Kit*
Cell Lysis Solution*
Nuclei Lysis Solution*
Protein Precipitation Solution*
DNA Rehydration Solution*
RNase A Solution, 4mg/ml*
Proteinase K*
Product
Wizard® SV Genomic
DNA Purification System(a)
Wizard® SV 96 Genomic
DNA Purification System(a)
Proteinase K*
Wizard® SV Lysis Buffer*
Wizard® SV Wash Solution*
Neclei Lysis Solution*
EDTA, 0.5M (pH 8.0),
Molecular Grade*
RNase A Solution, 4mg/ml*
Product
MagneSil® Blood Genomic,
Max Yield System(b)*
Anti-Foam Reagent*
MagneSil® Paramagnetic Particles*
Salt Wash, Blood*
Alcohol Wash, Blood*
Elution Buffer, Blood*
Product
MagneSil® ONE, Fixed Yield
Blood Genomic System(b)*
Lysis Buffer, Blood*
Anti-Foam Reagent*
MagneSil® PMPs—Fixed Yield*
Alcohol Wash, Blood*
Elution Buffer, Blood*
Size
Cat.#
100 isolations x 300µl
500 isolations x 300µl
100 isolations x 10ml
1 liter
50ml
1 liter
25ml
350ml
50ml
1ml
100mg
A1120
A1125
A1620
A7933
A7941
A7943
A7951
A7953
A7963
A7973
V3021
Size
Cat.#
50 preps
250 preps
A2360
A2361
1 x 96 preps
4 x 96 preps
100mg
50ml
185ml
50ml
A2370
A2371
V3021
Z3052
A1311
A7941
100ml
1ml
V4231
A7973
Size
Cat.#
1 x 96 preps
300µl
25ml
90ml
70ml
45ml
MD1360
MD1431
MD1441
MD1401
MD1411
MD1421
Size
Cat.#
1 x 96 preps
160ml
300µl
25ml
120ml
45ml
MD1370
MD1392
MD1431
MD1451
MD1412
MD1421
Product
Wizard® Magnetic 96 DNA
Plant System(b)
Wash Buffer, Plant
Product
MagneSil® Genomic, Fixed
Tissue System*
Product
MagneSil® KF, Genomic System*
MagneSil® KF, Paramagnetic Particles*
Lysis Buffer, KF*
Product
Wizard® Magnetic DNA Purification
System for Food(b)
RNase A Solution, 4mg/ml*
Lysis Buffer A, Food
Lysis Buffer B, Food
Precipitation Solution, Food
Hard Ware Devices/Accessories
Product
Vac-Man® Laboratory Vacuum Manifold,
20-sample capacity Vacuum Adapters
Vac-Man® 96 Vacuum Manifold
Deep Well MagnaBot® 96 Magnetic
Separation Device*
MagnaBot® Spacer 1/8 inch
Collection Plates (4-pack)
Heat Transfer Block
MagnaBot® 96 Magnetic
Separation Device
MagnaBot® Spacer
MagnaBot® Adapter T1
MagneSphere® Technology
Magnetic Separation Stand (two-position)
MagneSphere® Technology
Magnetic Separation Stand (twelve-position)
PolyATtract® System 1000
Magnetic Separation Stand
Size
Cat.#
2 x 96 preps
4 x 96 preps
40ml
FF3760
FF3761
A3811
Size
Cat.#
100 samples
MD1490
Size
200 preps
40ml
160ml
Cat.#
MD1460
MD1471
MD1521
Size
Cat.#
200 preps
400 preps
1ml
100ml
100ml
150ml
FF3750
FF3751
A7973
A8191
Z3191
Z3201
Size
1 each
20 each
1 each
Cat.#
A7231
A1331
A2291
1
1
1
1
each
each
each
each
V3031
V8581
A9161
Z3271
1 each
1 each
1 each
V8151
V8381
V8481
1.5ml
Z5332
1.5ml
Z5342
1 each
Z5410
Genomic DNA
Ordering Information
*For Laboratory Use.
(a)Australian
Pat. No. 730718 and other patents and patents pending.
Pat. Nos. 6,027,945, 6,368,800 and 6,673,631, Australian Pat. No. 732756 and other patents and
patents pending.
(b)U.S.
MagneBot, MagneSil, PolyATtract and Wizard are registered trademarks of Promega Corporation.
ABI PRISM is a registered trademark of Applera Corporation.
Biomek is a registered trademark of Beckman Coulter, Inc.
13
Information...always at your finger tips!
Learn how…ask Promega
DNA Analysis Notebook
Get the latest information to help you purify genomic DNA,
amplify and clone your targets.
RNA Analysis Notebook
Isolate RNA and analyze by RT-PCR, microarrays and even
RNAi.
Call your local Promega representative
to get your free copy or visit us on-line
at: www.promega.com/guides
#BR129
#BR120
#BR152
14
Purify Plasmid DNA...
...at high yield, on any scale from manual to automated
plate format.
Purify Plasmid on Any Scale:
• Miniprep
• Midiprep
Promega plasmid DNA purification systems provide:
• Your choice of purification method: Promega provides a range of
membrane filtration-based, magnetic particle-based and resin plasmid
DNA purification systems. You choose the separation method best
suited to your needs.
• Maxiprep
• Megaprep
• Multiwell-plate formats
• Full automation
Plasmid DNA
• Your choice of throughput: Both membrane-based and magnetic-based
plasmid purification systems are available in manual and automated
formats.
• Systems tailored to your purification scale: From miniprep to
megaprep scale, there is a plasmid purification system for every need.
• Fully automated systems for high-throughput users: Automated
methods and personalized automation support are available for all high
throughput systems.
• Increased productivity: Save time and labor with our easy to use
protocols.
How can we help you achieve your goals today?
A Comparison of Plasmid DNA Purification Systems.
Culture
Volume
PureYield™ Plasmid Midiprep System (Cat.# A2492, A2495)
Wizard® Plus
SV Minipreps (Cat.# A1330, A1340, A1460, A1470)
SpinProtocol
VacuumProtocol
Magnetic
Technology
Manual
Handling
Automated
Handling
50–250ml
1-10ml
Wizard® SV 96 Plasmid System (Cat.# A2250, A2255)
multiwell
Wizard® MagneSil Tfx™ System (Cat.# A2380, A2381)
multiwell
Wizard® Plus
Mini and Midipreps Systems (Cat.# A7100, A7500, A7510, A7640)
1-100ml
Wizard® Plus
Maxi and Megapreps Systems (Cat.# A7270, A7300)
100-1,000ml
Wizard® PureFection System (Cat.# A2150, A2160, A2170)
5-125ml
Wizard® MagneSil® Plasmid Purification System (Cat.# A1630, A1631)
multiwell
15
The PureYield™ Plasmid Midiprep System
Fast and reliable membrane-based column method for
midiprep plasmid DNA purification.
The PureYield Plasmid Midiprep System provides:
• Improve your productivity: Rapid protocol purifies large amounts
of plasmid DNA in 30 minutes.
• Have confidence in your results: Highly purified and concentrated
plasmid DNA gives proven performance in transfection in vitro
expression and other molecular biology applications.
• Spend less time purifying DNA: High yield gives you more
experiments per midiprep.
Size
Cat.#
25 preps
100 preps
A2492
A2495
For Research Use.
PureYield™ System Done in 30 Minutes
Wizard® System 99 Minutes
Qiagen Plasmid 200 Minutes
Comparison of time required per midiprep using different systems. Each system
protocol was performed according to the manufacturer’s instructions using 50ml of an overnight
culture of JM109 bacteria transformed with a high copy plasmid (pGEM®-3 plasmid). Total time
to perform the midiprep is noted.
Qiagen
Eppendorf PureYield™
System
DNA Preparation Methods
Comparison of transfection efficiency of purified plasmid DNA prepared using different
midiprep systems. Comparison of transfection of plasmid DNA purified with the PureYield
System and other midiprep systems. psiCHECK-2™ Vector (Cat.# C8021), which carries a firefly
luciferase gene, was isolated from E.coli using either the PureYield System, the HiSpeed™ Qiagen
Plasmid Midi Kit or the Eppendorf Perfectprep® Plasmid Midi Kit. HeLa cells were transfected
using 0.07µg of DNA in a total of 25µl. The firefly luciferase signal was monitored with the DualGloTM Luciferase Reporter System (Cat.# E1910).
4594CA
8,000
7,000
6,000
5,000
4,000
3,000
2,000
1,000
0
4701MA
Firefly Luciferase (RLU)
4587MB
• Flexibility: The high binding capacity of PureYield columns allows
plasmid purification from large culture volumes (250ml), providing
maxiprep yield from a midiprep system.
Product
PureYield™ Plasmid
Midiprep System
Proper assembly of Lysate Clearing Column (blue) and DNA Binding Column
(white) for use with the PureYield Plasmid Midiprep System vacuum-method
purification protocol.
16
Wizard® Plus SV Minipreps DNA Purification System
Low-throughput, silica-membrane column-based
miniprep plasmid DNA purification in manual format
The Wizard Plus SV Minipreps System provides:
• High yield: Up to 20µg of plasmid from 10ml culture.
• Sequencing-quality DNA: The system is quality tested in
fluorescent sequencing reactions to ensure read lengths of at least
500 base pairs with 98% accuracy.
Product
Wizard® Plus SV Minipreps DNA
Purification System
Wizard® Plus SV Minipreps DNA
Purification System + Vacuum Adapters
Wizard® Plus SV Minipreps DNA
Purification System
Wizard® Plus SV Minipreps DNA
Purification System + Vacuum Adapters
Size
Cat.#
50 preps
A1330
50 preps
A1340
250 preps
A1460
250 preps
A1470
For Laboratory Use.
(a) U.S. Pat. No. 4,766,072.
(b) U.S. Pat. No. 5,955,363.
Plasmid DNA
• Time savings: Isolate 20 minipreps in less than 45 minutes.
• Compatibility with EndA+ strains: Incorporation of an alkaline
protease step allows DNA isolation from problematic strains.
1612TA09_6A
1
2
3
4
5
6
M
kb
– 10.0
––––– 8.0
– 6.0
––––– 5.0
– 4.0
– 3.0
––––– 2.5
– 2.0
– 1.5
– 1.0
– 0.75
– 0.50
– 0.25
High-quality restriction digests using plasmid purified with the Wizard Plus SV
Minipreps DNA Purification System. High-copy pGEM®-3Zf(+) Vector(a) (lanes 1–3) and
low-copy pALTER®-1 Vector(a,b) (lanes 4–6) were each digested in two separate Promega
restriction enzyme reactions. All lanes show reproducible and efficient cutting of the plasmid DNA
and are measured against the 1kb DNA Ladder (Cat.# G5711).
pGEM® 3-Zf(+) plasmid sequenced with T7 sequencing primer using DNA
purified by the Wizard Plus SV Minipreps DNA Purification System.
17
Wizard® SV 96 Plasmid DNA Purification Systems
High-throughput 96-well silica-membrane plate based
plasmid DNA purification in automated or manual
format
Product
Wizard® SV 96 Plasmid
DNA Purification System
Wizard® SV 9600 Plasmid
DNA Purification System*
The Wizard SV 96 System provides:
• Sequencing-quality DNA: Quality tested in fluorescent sequencing
reactions to ensure read lengths of at least 600 base pairs with
98% accuracy.
• High quality at high speed: Process 96-wells of bacterial culture
in 60 minutes or less, using a 96-well, vacuum filtration step to
simultaneously clear bacterial cell lysates and bind DNA—no
centrifugation required!
Cat.#
1 x 96 preps
5 x 96 preps
A2250
A2255
100 x 96 preps
A2258
Cat.# A2250, A2255 For Laboratory Use.
*Cat.# A2258 is supplied with the components listed for Cat.# A2250 except for Alkaline Protease Solution,
Nuclease-Free Water, 96-Well Deep Well Plates, DNA Elution Plates and Plate Sealers.
Overnight cultures
Resuspend pelleted
bacterial cells.
Lyse.
Neutralize.
• A walk-away, automated procedure: No manual intervention
required once the E.coli cell pellets are placed on the robot deck.
• Hassle-free automated methods: Automated methods and
support information available at: www.promega.com/automethods/
Size
Transfer Lysates.
Lysate Clearing Plate
• Ease-of-use and handling: The novel plate design prevents cross
contamination during sample processing. Filtrate waste products
are delivered directly to a vacuum trap, eliminating the need to
empty waste collection vessels during the purification procedure.
Clear lysates.
Binding Plate
Bind DNA.
3758CA06_2A
Wash.
Elute DNA.
The Wizard® SV 96 System operates as a true “walkaway” system on automated purification
platforms such as the Beckman Coulter Biomek® FX laboratory automation workstation. It can
also be used in a manual format.
Wizard SV 96 is tested to ensure that purified
DNA meets or exceeds 600 bases with 98%
accuracy in fluorescent sequencing analysis.
Highly pure
plasmid DNA.
2651MB04_1A
Elution Plate
Wizard SV 96 Plasmid DNA Purification System protocol overview. Automated
protocols are available at: www.promega.com/tbs
18
Wizard MagneSil Tfx™ System
Transfection-quality plasmid DNA purification in a
high-throughput 96-well miniprep format
The Wizard MagneSil Tfx System provides:
• Lowered endotoxin plasmid DNA: Use of the endotoxin removal
resin cuts endotoxin carryover as much as 95% over automated
standard sequencing grade plasmid systems. Lower endotoxin
levels produce better transfection efficiency.
Product
Wizard MagneSil Tfx™ System
Size
4 x 96 preps
8 x 96 preps
Cat.#
A2380
A2381
Clear Cell Debris
• High-quality at high speed: Process 96-wells of bacterial culture
in under 45 minutes with a Beckman Coulter Biomek® FX, using a
paramagnetic particle-based purification method for lysate clearing
and DNA capture.
• Hassle-free automated methods: Automated method and support
information available at: www.promega.com/automethods/
Plasmid DNA
• Improved mammalian protein expression: Three- to five-fold
increase in protein expression compared to plasmid isolated from
an automated sequencing-grade plasmid purification system.
Remove Endotoxin
Bind DNA
Wash
Wash
20
15
10
5
0
3905MA12_2A
µg DNA
Yields of Plasmid DNA
1
48
96
Bind DNA
Dry
Sample-to-sample yield of plasmid DNA purified using the Wizard MagneSil Tfx System. Deepwell plates containing JM109 cells (4 O.D.600) transformed with pGL3 Control Vector DNA(a,b)
(Cat.# E1741) were processed by three different users following the standard automated protocol
provided in the Wizard MagneSil Tfx™ System Technical Bulletin #TB314. Each bar represents the
average of 8 samples. Samples were quantitated by gel electrophoresis.
Elute
4387MA11_3A
Sample Number
Flow chart of Transfection-grade plasmid DNA purification using the Wizard
MagneSil Tfx System.
(a)U.S.
Pat. No. 5,670,356.
method of recombinant expression of Coleoptera luciferase is covered by U.S. Pat. Nos. 5,503,024,
5,674,713 and 5,700,673. A license (from Promega for research reagent products and from the Regents of the
University of California for all other fields) is needed for any commercial sale of nucleic acid contined within or
derived from this product.
(b)The
19
Wizard® Plus DNA Purification Systems
Mini-, Midi-, Maxi- and Megaprep scale plasmid DNA
purification in manual format
The Wizard Plus Systems provide:
• Simplicity: Only a centrifuge, a vacuum source and a vacuum
manifold required.
• Fast, manual format: Minipreps can be completed in as little as
15 minutes, Midipreps in 90 minutes, and Maxi- and Megapreps
in 3 hours.
Product
Wizard® Plus Minipreps DNA
Purification System
Wizard® Plus Midipreps
DNA Purification System
Wizard® Plus Maxipreps
DNA Purification System
Wizard® Plus Megapreps
DNA Purification System
Size
Cat.#
50 preps
100 preps
250 preps
A7100
A7500
A7510
25 preps
A7640
10 preps
A7270
5 preps
A7300
• Sequencing-quality DNA: Each Wizard Plus System is quality
tested in fluorescent sequencing reactions to ensure read lengths
of at least 500 base pairs with 98% accuracy.
• Safety and convenience: No phenol extraction or precipitation
required.
• High yields: Using a high copy number plasmid, the following
yields are obtained: 1–10µg DNA from 1–3ml culture; up to 200µg
DNA from 10–100ml culture; up to 300µg–1mg from 100–500ml
culture; and >1mg from 1,000ml culture.
Fluorescent dye primer sequencing of pGEM® Vector purified using the Wizard Plus
Minipreps DNA Purification System. The system is tested to ensure that purified
plasmid DNA can be sequenced at ≥98% accuracy over 500 bases using dye primer
chemistry and an ABI PRISM® 377 DNA sequencer.
High Copy Number
Low Copy Number
200
150
100
1460MA04_6A
DNA purification using the Wizard Plus
Systems requires use of a vacuum source
and either a lab-assembled or commercial
manifold such as Promega’s Vac-Man ®
Laboratory Vacuum Manifold (20-sample
capacity).
Plasmid Yield (µg)
250
50
0
25
50
75
100
Culture Volume (ml)
(a)U.S.
20
Pat. No. 4,766,072.
Plasmid DNA yield obtained using the Wizard Plus Midipreps DNA Purification
System. Yield is shown as a function of culture volume and plasmid copy number.
Representative yields of a high-copy number plasmid, pGEM®-3Zf(+) Vector(a), and a
low-copy plasmid, pBR322, prepared in E. coli strain DH5α™.
Wizard® PureFection Plasmid DNA Purification System
The Wizard PureFection System provides:
• Lower endotoxin plasmid DNA: Use of the endotoxin removal resin
cuts endotoxin carryover as much as 95% over automated standard
sequencing grade plasmid systems. Lower endotoxin levels produce
better transfection efficiency.
• Safety and convenience: No phenol extraction or precipitation
required, using a magnetic-based purification method for lysate
clearing and DNA capture.
Product
Size
Wizard® PureFection Plasmid
DNA Purification System
2 maxipreps/10 midipreps
5 maxipreps/25 midipreps
25 maxipreps/125 midipreps
Cat.#
A2150
A2160
A2170
For Laboratory Use.
Alkaline Lysis
Produce cleared lysate.
Plasmid DNA
Low-throughput, transfection-grade plasmid DNA
purification in scaleable manual format
• High yield: 40µg DNA from 5ml culture; scale-up capacity up to
~1mg.
• Fast, efficient purification: The hand-held MagneSil Magnetic
Separation Unit holds a 50ml tube and allows rapid and efficient
capture of magnetic particles and their attached DNA.
Selective Removal of
Endotoxin with Magnet
Endotoxin Removal Resin
clears bacterial lysate of
endotoxin.
• Scalable format: Rapid purification from a range of starting volumes
from midiprep (5–25ml) to maxiprep (26–125ml) scale.
Bind Plasmid DNA
700
Selective binding of plasmid
DNA using MagneSil®
Paramagnetic Particles.
600
400
Wash & Elute
Wash particles to remove
salts and elute in water.
Ethanol precipitate and
resuspend DNA at
desired concentration.
300
200
2347MB08_8A
Endotoxin (EU/ml x 103)
500
100
0
0
1
2
3
4
5
6
7
Endotoxin Removal Resin (mg)
8
9
10
2351MA08_8B
Overview of Wizard PureFection Plasmid DNA Purification System protocol.
Removal of endotoxin from bacterial cell lysates using Endotoxin Removal Resin. Over 95% of
endotoxin is removed from bacterial lysates following a 5-minute incubation.
21
Wizard® MagneSil® Plasmid Purification System
Product
Wizard® MagneSil® Plasmid
Purification System
The Wizard MagneSil Plasmid Purification System provides:
• A walk-away automated procedure: No manual intervention
required once e. coli cell pellets are placed on robot deck.
• High-throughput: Process multiple plates without user
intervention.
• A simple, fast method: Plasmid DNA isolation is accomplished
easily, with a simple, magnetic-based procedure, using a
paramagnetic particle-based purification method for lysate clearing
and DNA capture.
Cat.#
4 x 96 preps
8 x 96 preps
100 x 96 preps
A1630
A1631
A1635
12
Average Yield (µg)
• Sequencing-quality DNA: The system is quality tested in
fluorescent sequencing reactions to ensure read lengths of at least
600 base pairs with 98% accuracy.
Size
• Hassle-free automated methods: Automated methods and
support information available at: www.promega.com/automethods/
11
10
9
8
7
6
5
4
3
2
1
0
2
4
8
6
Biomass O.D.600
The Wizard MagneSil Plasmid Purification System is tested to ensure reliable DNA
recovery. Plasmid DNA was purified from multiple samples grown to the stated
O.D.600. DNA yield was measured on an agarose gel.
Capture MagneSil® BLUE
on MagnaBot® Device.
Clear Cell Debris
Wash
Capture the MagneSil® RED
on the MagnaBot® Device.
Dry
Wizard MagneSil Plasmid Purification System protocol. Automated protocols are available at:
www.promega.com/tbs
Accuracy (%)
Elute
2 O.D.
4 O.D.
6 O.D.
8 O.D.
10 O.D.
12 O.D.
100
4388MA
Remove supernatant.
Bind DNA
3193MB06_1A
High-throughput 96-well plasmid DNA purification in
an automated format
98
96
94
90
MagneSil Paramagnetic Particles can be
relied upon to purify high-quality template
for fluorescent sequencing.
22
500
600
Bases
700
2890MA03_0A
92
Accuracy by read length for plasmid DNA sequenced following isolation by
MagneSil Paramagnetic Particles. Results demonstrate greater than 700 consecutive
bases analyzed with greater than 98% accuracy of base identity.
Ordering Information
Size
25 preps
100 preps
Cat.#
A2492
A2495
Size
Cat.#
50 preps
A1330
50 preps
A1340
250 preps
A1460
250 preps
3ml
A1470
A1441
Size
Cat.#
1 x 96 preps
5 x 96 preps
A2250
A2255
100 x 96 preps
185ml
370ml
500ml
950ml
Wizard® SV 96 Cell Resuspension Solution*
500ml
800ml
Wizard® SV 96 Cell Lysis Solution*
500ml
800ml
Nuclease-Free Water*
150ml
Alkaline Protease Solution*
3ml
A2258
A1311
A1318
A1481
A1488
A7113
A7118
A7123
A7128
P1195
A1441
Product
Wizard MagneSil Tfx™ System(a,d,e)
Size
4 x 96 preps
8 x 96 preps
100ml
115ml
Cat.#
A2380
A2381
A2191
A2221
Size
Cat.#
Product
Wizard® Plus SV Minipreps DNA
Purification System(b,c)*
Wizard® Plus SV Minipreps DNA
Purification System + Vacuum Adapters(b,c)*
Wizard® Plus SV Minipreps DNA
Purification System(b,c)*
Wizard® Plus SV Minipreps DNA
Purification System + Vacuum Adapters(b,c)*
Alkaline Protease Solution*
Product
Wizard® SV 96 Plasmid DNA
Purification System(b)*
Wizard® SV 9600 Plasmid DNA
Purification System
Wizard® SV Wash Solution*
Wizard® SV 96 Wash Solution
Wizard® SV 96 Neutralization Solution*
Endotoxin Removal Resin
4/40 Wash Solution
Wizard® Plus Megapreps DNA
Purification System(f)*
Product
Wizard® PureFection Plasmid DNA
Purification System(a,d)*
Product
Wizard® MangeSil® Plasmid
Purification System(d,e)
MagneSil® RED
MagneSil® BLUE
Cell Resuspension Solution
Cell Lysis Solution
Neutralization Solution
Elution Buffer
5 preps
A7300
Size
Cat.#
2 maxipreps/
10 midipreps
5 maxipreps/
25 midipreps
25 maxipreps/
125 midipreps
A2150
A2160
A2170
Size
Cat.#
4 x 96 preps
8 x 96 preps
100 x 96 preps
100ml
100ml
500ml
500ml
500ml
500ml
A1630
A1631
A1635
A1641
A2201
A7114
A7124
A7132
A1655
Size
Cat.#
1 each
A7231
Plasmid DNA
Product
PureYield™ Plasmid Midiprep System(a)
Hard Ware Devices/Accessories
Product
Vac-Man® Laboratory Vacuum Manifold,
20-sample capacity
Vac-Man® Jr. Laboratory Vacuum Manifold,
2-sample capacity
Vacuum Adapters
Vac-man 96 Vacuum Manifold
Collar for Vac-Man 96 Vacuum Manifold
Wizard® SV 96 Binding Plates*
Wizard® SV 96 Lysis Clearing Plates
MagnaBot® 96 Magnetic Separation Device
Collection Plates (4-pack)
1
20
1
1
10
100
10
100
1
1
each
each
each
each
pack
each
pack
each
each
each
A7660
A1331
A2291
A2311
A2271
A2278
A2241
A2248
V8151
A9161
*For Laboratory Use.
Product
Wizard® Plus Minipreps DNA
Purification System(f)*
Wizard® Plus Midipreps DNA
Purification System(f)*
Product
Wizard® Plus Maxipreps DNA
Purification System(f)*
(a)U.S.
50 preps
100 preps
250 preps
A7100
A7500
A7510
25 preps
A7640
Size
Cat.#
10 preps
A7270
Pat. No. 6,194,562, Australian Pat. No. 740145, Canadian Pat. No. 2,329,067 and other patents
pending.
(b)U.S. Pat. No. 5,981,235, Australian Pat. No. 729932 and other patents pending.
(c)Australian Pat. No. 730718 and other patents and patents pending.
(d)U.S. Pat. Nos. 6,027,945, 6,368,800 and 6,673,631, Australian Pat. No. 732756 and other patents and
patents pending.
(e)U.S. Pat. No. 6,284,470 and other patents pending.
(f)U.S. Pat. Nos. 5,658,548 and 5,808,041, Australian Pat. No. 689815 and other patents and patents
pending.
MagneSil, pALTER, pGEM, Vac-Man and Wizard are registered trademarks of Promega Corporation.
23
Superior Technical Services
Scientists helping scientists
to be successful!
Get the best-in-class help and support you need from Promega.
All Promega Technical Services Scientists have extensive
laboratory experience. All have life science degrees and most
have post-graduate or post-doc lab experience. If you have a
question about one of our products, or even about a product
that is not ours, feel free to contact us.
www.promega.com
Visit our web site for 24-hour a day support. Find online protocols, product
FAQs, application information, references, publications, calculators and many
other valuable support tools. It’s all easy to access and easy to understand.
■
Telephone
■
E-mail
■
Troubleshooting
experiments
■
Automation installation
and support
■
Global availability
■
Multi-language capabilities
Telephone: 608-274-4330
Toll Free in USA: 800-356-5926
www.promega.com/techserv/
24
Purify DNA Fragments...
...from PCR, Sequencing Reactions and Gel Slices.
Promega fragment DNA purification systems provide:
Purify Fragments:
• Directly from PCR
• From gels
• High yield: Excellent recovery rates direct from PCR or from gels.
• From sequencing reactions
• Your choice of purification method: We provide a range of membrane
filtration-based and magnetic particle-based systems. You choose the
separation method best suited to your needs.
• From restriction digests
• Away from phosphatases, kinases,
polymerases
• Your choice of throughput: Both membrane-based and magnetic-based
purification systems are available in manual and automated formats.
• Simplicity and sophistication: From manual, single-tube purification to
full automation, there is a fragment purification system for every need.
• Fully automated systems for high-throughput users: Automated
methods and personalized automation support are available for all high
throughput systems.
Fragment DNA
• Increased productivity: Save time and labor from easy to use
protocols.
• Experience you can trust: Have confidence in a supplier with over 25
years experience providing molecular biology systems and reagents.
How can we help you achieve your goals today?
A Comparison of Fragment DNA Purification Systems.
Sequencing
Clean-Up
PCR
Gel Fragment
Clean-Up
Isolation
Spin
Protocol
Vacuum
Protocol
Magnetic
Technology
Manual
Handling
Automated
Handling
Wizard® SV Gel and PCR Clean-Up (Cat.# A9281, A9282)
Wizard® SV 96 PCR Clean-Up (Cat.# A9340, A9341, A9342)
Wizard® PCR Preps (Cat.# A7170, A2180)
Wizard® DNA Clean-Up (Cat.# A7280)
Wizard® MagneSil® Sequencing Reaction Clean-Up
(Cat.# A1830, A1831, A1832)
Wizard® MagneSil® PCR Clean-Up (Cat.# A1930, A1931)
25
Wizard® SV Gel and PCR Clean-Up System
High recovery of concentrated DNA fragments over a
broad size range.
Product
Wizard® SV Gel and PCR
Clean-Up System
The Wizard SV Gel and PCR Clean-Up System provides:
• Speed: Purify DNA fragments or PCR products in as little as 20
minutes directly from PCR or from agarose gels in either TAE or
TBE buffer.
For Laboratory Use.
• High yield: 70–95% recovery of DNA fragments (100bp–10kb)
from gels and 85–95% recovery of PCR products (100bp–3.2kb).
Recovery of PCR products >3.2kb is similar to that from agarose
gels.
M
100bp
U
P
500bp
U
P
Size
Cat.#
50 preps
250 preps
A9281
A9282
1,000bp
U
P
3,199bp
U
P
3,000bp –
1,000bp –
• Safety and convenience: No phenol extraction or precipitation
required.
Gel Purification
Excise gel slice
and add equal
volume of Membrane
Binding Solution
Direct PCR Product
Purification
50–65°C for 10
minutes to solubilize
gel slice
Add equal volume
of Membrane Binding
Solution to PCR reaction
Centrifuge
Vacuum
Adapter
(Cat.# A1331)
Bind DNA
U: Unpurified
P: Purified
Gel analysis of DNA fragments before and after gel extraction using the Wizard SV
Gel and PCR Clean-Up System. DNA fragments of the sizes indicated were analyzed
before (U; unpurified) and after (P; Purified) purification. The marker (M) used is the
1kb DNA Ladder (Cat.# G5711).
Wizard SV Gel and PCR Clean-Up System: Percent recovery of various
sized fragments from agarose gels.
Transfer sample to SV Minicolumn
Vacuum
3789TA07_2A
• An application-tested system: Tested to ensure performance of
purified PCR products in fluorescent sequencing. Purified DNA
routinely achieves 700bp with >98% accuracy.
DNA Fragment Size
Percent Recovery
70bp
85bp
100bp
500bp
1,000bp
3,199bp
9,416bp
39%
55%
84%
89%
92%
95%
95%
Wizard SV Gel and PCR Clean-Up System: Percent recovery of various
sized fragments directly from PCR.
Wash, removing
solution by
centrifugation
or vacuum
Transfer spin
column to a 1.5ml
microcentrifuge
tube (not provided)
Centrifuge
Elute DNA fragment
or PCR product
3792MA07_2A
Vac-Man®
Laboratory
Vacuum Manifold
(Cat.# A7231)
Flow chart of DNA fragment gel purification or direct PCR product purification using the
Wizard SV Gel and PCR Clean-Up System.
26
DNA Fragment Size
Percent Recovery
200bp
500bp
1,000bp
1,500bp
3,199bp
87%
91%
92%
92%
87%
Wizard® SV 96 PCR Clean-Up System
Rapid high-throughput 96-well recovery of
fragment DNA
Product
Wizard® SV 96 PCR
Clean-Up System
The Wizard SV 96 PCR Clean-Up System provides:
• Speed: Process a 96-well plate in 20 minutes.
For Laboratory Use.
• High yield: Average percent recovery for 500bp and 1,000bp
fragments is >90%, unaffected by common PCR additives.
1
4
8
100
x
x
x
x
96
96
96
96
Size
Cat.#
preps
preps
preps
preps
A9340
A9341
A9342
A9345
A.
• Gel slice purification: The system can be used to purify DNA from
gel slices, as well as directly from PCR.
M
100bp
200bp
300bp
500bp
P
P
P
P
U
U
U
U
1,000bp
P
U
• Safety and convenience: No phenol extraction or precipitation
required.
3801TA08_2A
• An application-tested system: Tested to ensure performance of
purified PCR products in fluorescent sequencing and microarray
applications.
• Hassle-free automated methods: Automated methods and
support information available at: www.promega.com/automethods/
Percent Recovery
Mix PCR sample and Membrane
Binding Solution in equal volumes.
Apply to Binding Plate.
Apply vacuum to bind
PCR product to membrane.
100
90
80
70
60
50
40
30
20
10
0
100
300
Size (bp)
Wash.
3743MA06_2A
Collection Plate
Elute bound PCR product with
Nuclease-Free Water.
200
500
1,000
3801TB08_2A
Binding Plate
Fragment DNA
B.
Purification and recovery of PCR products. PCR fragments of 100, 200, 300, 500, and
1,000 base pairs were purified using the Wizard SV 96 PCR Clean-Up System on the
Beckman Coulter Biomek® 2000 robotic workstation. Panel A. Purified (P) and
unpurified (U) fragments were separated on an ethidium bromide-stained, 2% agarose
gel. Panel B. Percent recovery of purified PCR products was quantitated using a
Hitachi FMBIO® Fluorescent Scanner. Results show the mean and standard deviation
for 6 purified fragments of each size. For small PCR fragments (<500bp), optimal
recovery requires a 95% ethanol wash. For larger fragments (>500bp), optimal results
are achieved using an 80% ethanol wash.
Wizard SV 96 PCR Clean-Up System protocol overview.
27
Wizard® PCR Preps DNA Purification System
Reliable low-throughput PCR and agarose gel slice
DNA fragment clean-up
Product
Wizard® PCR Preps
DNA Purification System
The Wizard PCR Preps DNA Purification System provides:
• High yield: 70–90% recovery of DNA fragments (500bp) from
low-melting point agarose gels and 95–99% recovery of PCR
products (300bp–1.5kb) directly from reactions.
For Laboratory Use.
1
2
3
4
Size
Cat.#
50 preps
250 preps
A7170
A2180
5
• Efficiency: Process multiple samples simultaneously using the
Vac-Man Laboratory Vacuum Manifold (20 sample capacity).
• Safety and convenience: No phenol extraction or precipitation
required.
Before Purification
M
0348TA08_3A
• Confidence: Removal of excess nucleotides, primers and small
nonspecific amplification products such as primer-dimers from
PCR.
After Purification
M
Purification and analysis of PCR products from low-melting agarose using the
Wizard PCR Preps DNA Purification System. Lane 1, 250ng pGEM® DNA Markers;
lane 2, β-1-anti-trypsin PCR product; lane 3, purified β-1-antitrypsin PCR fragment;
lane 4, b-globin PCR reaction product; lane 5, purified b-globin PCR fragment.
1,000 –
500 –
200 –
50 –––––
0935TA11_4A
bp
Recovery of PCR products using the Wizard PCR Preps Resin. A representative sample from the
simultaneous purification of 96 PCR reactions was chosen to determine the effectiveness of the
DNA purification process. Equivalent amounts from before and after purification were separated on
a 1% agarose gel and stained with ethidium bromide.
28
Wizard® DNA Clean-Up System
Safe, easy clean-up of linear or circular DNA
Product
Wizard® DNA Clean-Up System
Size
100 preps
Cat.#
A7280
For Laboratory Use.
The Wizard DNA Clean-Up System provides:
• Speed: Purify DNA in as little as 15 minutes.
• Efficiency: Process multiple samples simultaneously using the
Vac-Man Laboratory Vacuum Manifold (20–sample capacity).
• Safety and convenience: No phenol extraction or precipitation
required.
• Applications tested: Eluted DNA demonstrated for use in cloning,
clean-up of hi-sulfate treated genomic DNA and other applications.
Removal of Common Impurities by the Wizard DNA Clean-Up System.
Contaminant
Restriction enzymes
Heat-stable restriction enzymes
Phosphatases
Kinases
Ligases
DNA polymerases
Taq DNA polymerases
Exonucleases
DNase I
RNases
Mononucleotides
Salts
Oligonucleotides
Removed
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
No
Yes
Yes
No
Fragment DNA
• Simple, effective purification: Purify linear and circular DNA
(200bp–50kb) away from contaminants such as restriction
enzymes, phosphatases, kinases, DNA polymerases, exonucleases
and endonucleases (including DNase I but not RNases),
mononucleotides and salts.
29
Wizard® MagneSil® Sequencing Reaction Clean-Up System
High-throughput automated 96-well or 384-well
sequencing reaction clean-up made simple and
economical
The Wizard MagneSil Plasmid Purification System provides:
• High-throughput: Process multiple 96-well or 384-well plates
without user intervention.
• A simple, fast method: DNA isolation is accomplished easily, with
a simple, magnetic-based procedure.
Product
Size
Cat.#
preps
preps
preps
preps
A1830
A1831
A1832
A1835
Sequencing Reaction Clean-Up in 384-Well Plates
MagneSil® GREEN
100ml
MagnaBot® 384 Magnetic Separation Device 1 each
384-Well Plate, Conical
10/pack
A8231
V8241
V5311
Sequencing Reaction Clean-Up in 96-Well Plates
Wizard® MagneSil® Sequencing
Reaction Clean-Up System
1 x 96
4 x 96
8 x 96
100 x 96
Cat.# A1830, A1831, A1832, A1835, A8231, V8241 For Laboratory Use.
• High quality purified DNA: Typical Phred 20 values of 600–700
bases. 98% accuracy to at least 600 bases.
• Efficiency: Save labor with a walk-away high-throughput
sequencing reaction clean-up automated method.
3312CA_1A
• Hassle-free automated methods: Automated methods and
support information available at: www.promega.com/automethods/
35
# of Samples
30
Clean-up is performed using the MagnaBot II Magnetic Separation Device (Cat.#
V8351) accompanied by the Plate Clamp 96 (Cat.# V8251). These devices are
designed to work with most robotic platforms.
25
20
15
10
5
Phred 20 values for sequencing data from each well of a 96-well plate using the Wizard
MagneSil Sequencing Reaction Clean-Up System. Phred is a base-calling program that assigns
an error probability to each base. A phred value of 20 for a given base assigns an error probability
of 1 in a 100 for that base.
3995CA02_3A
Phred 20 Read Length (bp)
3271MA02_1B
70
0
72
5–
67
5
69
9–
65
0
67
4–
62
5
9–
64
62
4–
60
0
0
MagnaBot 384 Magnetic Separation Device (Cat.# V8241). The MagnaBot 384 is
constructed such that the magnetic field draws particles to the corners of the
processing plate.
Wizard MagneSil Sequencing Reaction
Clean-Up System undergoes rigorous testing
for performance. DNA prepared
by this method routinely achieves
98% accuracy out to 600 bases.
30
Sequencing
Reaction
Add Ethanol
Wash Solution
Add Sequencing
Gel Loading Solution
Place on
MagnaBot® II
Place on
MagnaBot® II
Place on
MagnaBot® II
Transfer eluate
to 96-well plate for
sequence analysis
Binding
Washing
Elution
Fragment DNA
Remove
Supernatant
3169MB04_1A
Remove
Supernatant
Add
MagneSil® GREEN
Overview of the Wizard MagneSil Sequencing Reaction Clean-Up System protocol.
A. MagneSil® PMPs
3250TA02_1A
B. EtOH Precipitation
A comparison of electropherograms of purified sequencing reactions analyzed on an ABI PRISM® 3700 DNA Sequencer. Panel A: Reactions purified with
the Wizard MagneSil Sequencing Reaction Clean-Up System. Panel B: Reactions purified by ethanol precipitation.
31
Wizard® MagneSil® PCR Clean-Up System
High-throughput PCR clean-up ideal for 96-well or
384-well automated format
The Wizard MagneSil PCR Clean-Up System provides:
• High-throughput: Process multiple 96-well or 384-well plates
without user intervention using a magnetic-based purification
method for DNA capture that is ideally suited for automation.
• A simple, fast method: No centrifugation or vacuum manifold
required.
• High quality DNA: Primer removal is consistently >98% with
purification of PCR fragments from 150bp–23kb in size.
PCR Clean-Up in 96-Well Plates
Product
Wizard® MagneSil®
PCR Clean-Up System(a)
Size
Cat.#
4 x 96 preps
8 x 96 preps
100 x 96 preps
A1930
A1931
A1935
Product
Size
MagneSil® YELLOW
100ml
Wash Solution
500ml
MagnaBot® 384 Magnetic Separation Device 1 each
384-Well Plate, Flat
10/pack
Cat.#
A9231
A8241
V8241
V5291
PCR Clean-Up in 384-Well Plates
Cat.# A1930, A1931, A1935, A8241, A9231, V8241 For Laboratory Use.
• An application-tested system: Tested to ensure performance of
purified PCR products in fluorescent sequencing and microarray
applications.
• Hassle-free automated methods: Automated methods and
support information available at: www.promega.com/automethods/
3993CA02_3A
100
% Recovery
80
60
MagnaBot 96 Magnetic Separation Device (Cat.# V8151).
40
20
Recovery of PCR products of varying size using the Wizard MagneSil PCR Clean-Up System. A
range of sizes of PCR products, generated using PCR Master Mix(a) (Cat.# M7505), were recovered
using the Wizard MagneSil PCR Clean-Up System on a liquid-handler. The yield of each PCR
product was determined by electrophoresis on a 2% agarose gel stained with ethidum bromide.
The mean and standard deviation of four separate determinations for each PCR product are
shown.
(a)
The PCR process is covered by patents issued and applicable in certain countries*. Promega does not encourage or
support the unauthorized or unlicensed use of the PCR process.
*In the U.S., effective March 29, 2005, U.S. Pat. Nos. 4,683,195, 4,965,188 and 4,683,202 will expire. In Europe, effective
March 28, 2006, European Pat. Nos. 201,184 and 200,362 will expire.
32
3995CA02_3A
50
0
3803MA08_2A
PCR Product Size (bp)
1,
00
0
1,
70
0
60
0
0
50
0
0
30
20
10
0
0
MagnaBot 384 Magnetic Separation Device (Cat.# V8241). The MagnaBot 384 is
constructed such that magnetic field draws the particles to the corners of the wells of
the processing plate.
Add
Wash Solution
PCR
Reactions
Add
Nuclease-Free
Water
Place on
MagnaBot® 96
Device
Add
MagneSil® YELLOW
Place on
MagnaBot® 96
Device
Place on
MagnaBot® 96
Device
Remove
Supernatant
DNA Binding
Washing
Elution of DNA
3288MB03_1A
Repeat wash
cycle 2X with
80% ethanol.
Fragment DNA
Transfer eluate
to 96-well plate.
Remove
Supernatant
Overview of the Wizard MagneSil PCR Clean-Up System protocol.
10
M
0
50 %
%
25
%
12
.5
6. %
25
%
Unpurified
Purified
bp
1,500 –
100 –
3493TA07_1A
500 –
The Wizard MagneSil PCR Clean-Up System removes primer-dimers from PCR products with
high yield and consistent results. A titration of unpurified prothrombin PCR product is shown
(100%, 50%, 25%, 12.5% and 6.25% of the PCR sample). The ~100bp primer-dimers are visible
in these lanes. Eight replicates purified using the Wizard MagneSil PCR Clean-Up System show the
506bp prothrombin PCR product with no visible primer-dimers. The ethidium bromide-stained 1%
agarose gel was imaged using an Alpha Innotech FluorChem™ fluorescence imaging system. Lane
M, 100bp DNA Ladder (Cat.# G2101).
33
Ordering Information
Product
Wizard® SV Gel and PCR Clean-Up System*
Size
preps
preps
preps
preps
preps
preps
20ml
Cat.#
A9281
A9282
A9340
A9341
A9342
A9345
A9301
Product
Wizard® PCR Preps DNA Purification System(b)*
Size
50 preps
250 preps
Cat.#
A7170
A2180
Product
Wizard® DNA Clean-Up System(c)*
Size
100 preps
Cat.#
A7280
Size
Cat.#
preps
preps
preps
preps
100ml
A1830
A1831
A1832
A1835
A8231
Size
Cat.#
4 x 96 preps
8 x 96 preps
100 x 96 preps
100ml
500ml
150ml
A1930
A1931
A1935
A9231
A8241
P1195
Wizard® SV 96 PCR Clean-Up System(a)*
1
4
8
100
50
250
x 96
x 96
x 96
x 96
Membrane Binding Solution*
Product
Wizard® MagneSil® Sequencing
Reaction Clean-Up System(d)*
MagneSil® GREEN*
Product
Wizard® MagneSil® PCR
Clean-Up System(d)*
MagneSil® YELLOW*
Wash Solution*
Nuclease-Free Water*
34
1
4
8
100
x
x
x
x
96
96
96
96
Hard Ware Devices/Accessories
Product
Vac-Man® 96 Vacuum Manifold
Vacuum Adapters
Wizard® SV 96 Binding Plates*
MagnaBot® II Magnetic Separation Device
Plate Clamp 96
Plate Stand
MagnaBot® 96 Magnetic Separation Device
Plate Clamp 96
Plate Stand
MagnaBot® Spacer
MagnaBot® Adapter T1
Collection Plates (4-pack)
MagnaBot® 384 Magnetic Separation Device*
384-Well Plate, Conical
384-Well Plate, Flat
1
20
10
100
1
1
1
1
1
1
1
1
1
1
10
10
Size
each
each
pack
each
each
each
each
each
each
each
each
each
each
each
pack
pack
Cat.#
A2291
A1331
A2271
A2278
V8351
V8251
V8261
V8151
V8251
V8261
V8381
V8481
A9161
V8241
V5311
V5291
*For Laboratory Use.
(a)Australian
Pat. No. 730718 and other patents and patents pending.
under U.S. Pat. No. 5,075,430.
(c)U.S. Pat. Nos. 5,658,548 and 5,808,041, Australian Pat. No. 689815 and other patents and patents
pending.
(d)U.S. Pat. Nos. 6,027,945 and 6,368,800, Australian Pat. No. 732756 and other patents and patents
pending.
(b)Licensed
MagnaBot, MagneSil, pGEM and Wizard are registered trademarks of Promega Corporation.
Dual-Glo is a trademark of Promega Corporation.
Biomek is a registered trademark of Beckman Coulter, Inc.
FMBIO is a registered trademark of Hitachi Software Engineering Company, Ltd.
ABI PRISM is a registered trademark of Applera Corporation.
FluorChem is a registered trademark of Alpha Innotech Corporation.
HiSpeed is a trademark of Qiagen.
Perfectprep is a registered trademark of Eppendorf.
Isolate RNA...
...from a variety of sample materials, on a scale of your
choosing.
Purify RNA From:
• Blood
• Tissue
Promega’s Total RNA and mRNA Isolation Systems provide:
• Cultured Cells
• Plant Material
• Your choice of purification method: Promega provides a range of
solution, membrane filtration-based and magnetic particle-based
systems. You choose the separation method best suited to your needs.
• Bacteria
• Your choice of throughput: Both membrane-based and magnetic-based
RNA isolation systems are available in manual and automated formats.
• Systems tailored to your purification scale: Mini-isolation from small
sample sizes to scaleable, solutions-based formats, there is a total RNA
purification system for every need.
• Direct isolation of total RNA or mRNA from biological materials:
Systems are available for direct isolation from a variety of starting
materials.
• Increased productivity: Save time and labor with our easy to use
protocols.
How can we help you achieve your goals today?
Total
RNA
Blood
Bacteria
Animal
Tissue
Culture
Cells
Plant
Material
Manual Automated
Handling Handling
Total RNA
A Comparison of RNA Isolation Systems.
Magnetic Spin
Vacuum
Technology Protocol Protocol
Total RNA Isolation Systems
RNAgents® Total RNA Isolation System (Cat.# Z5110)
SV Total RNA Isolation System (Cat.# Z3100, Z3105)
SV 96 Total RNA Isolation System (Cat.# Z3500, Z3505)
MagneSil® Total RNA mini-Isolation System (Cat.# Z3351)
mRNA Isolation Systems
PolyATtract® Automated System (Cat.# Z5671)
PolyATtract® System 1000 (Cat.# Z5400, Z5420)
PolyATtract® mRNA Isolation Systems (Cat.# Z5200, Z5300)
35
RNAgents® Total RNA Isolation System
Scalable solution-based total RNA purification for
maximum flexibility
Product
RNAgents® Total RNA Isolation System
The RNAgents Total RNA Isolation System provides:
• Speed: The solution-based procedure can be completed in 90
minutes to 3 hours.
Average Yields of Total RNA from a Variety of Tissues.
• Scalable: Isolate RNA from as little as 5mg, as much as 6g of
tissue, or from 105–108 cultured cells.
• Simplicity: A solution-based protocol for RNA isolation that can
be performed with ease in any laboratory setting.
• Performance: Yield and purity for today’s demanding applications
(e.g. microarrays and qRT-PCR)
• Complete system: Purity total RNA from blood, tissue cells, plant
leaf, etc.
Size
Scalable
Cat.#
Z5110
For Laboratory Use.
Tissue
RNAgents Total RNA Yield
HeLa cells
Human white blood cells
Mouse intestine
Mouse spleen
Mouse lung
Mouse kidney
Mouse liver
1.6mg RNA/108 cells
1.3mg RNA/108 cells
2.3mg RNA/g tissue
8.3mg RNA/g tissue
1.9mg RNA/g tissue
3.1mg RNA/g tissue
6.6mg RNA/g tissue
E. coli Control Array
1
2
3
4
5
6
Cy®3
Cy®5
3578TA11_1A
• Maiximum flexibility: Denaturing Solution available separately for
maximum flexibility, allowing you the option to supply the other
system components (phenol/chloroform, isoamyl alcohol and
isopropanol).
B. Jurkat Control Array
Cy®3: β-actin
Cy®5: Kan
– 28S
0349TA11_5A
– 18S
Analysis of RNA isolated from various tissues with the RNAgents System. RNA was isolated
from the following sources: HeLa cells (lane 1), mouse intestine (lane 2), mouse spleen (lane 3),
mouse lung (lane 4), mouse kidney (lane 5) and mouse liver (lane 6). Five micrograms of each
sample was resolved on a 1% denaturing agarose gel (2.2M formaldehyde).
36
Hybridization of fluorescent cDNA targets to E.Coli and Jurkat Cell Arrays. Panel A:
Hybridization results for the E. coli control array. One array was hybridized with a Cy®3
target and another with a Cy®5 target. Panel B: Hybridization results for the Jurkat
Control Array. Fluorescent Cy®3 and Cy®5 targets were co-hybridized to the same
array, demonstrating specificity for their complementary sequences.
SV Total RNA Isolation System
Simple and reliable total RNA purification from
tissues, cultured cells or white blood cells
Product
SV Total RNA Isolation System
The SV Total RNA Isolation System provides:
For Laboratory Use.
• Speed: The procedure can be completed in 60 minutes.
• Confidence in your results: Includes a DNase I treatment step to
ensure removal of genomic DNA. No detectable DNase I carryover
to the final RNA preparation.
• Peace of mind: Only system that includes DNase treatment step in
the standard protocol.
• Applications tested RNA: RNA suitable for all routine molecular
biology applications, including RT-PCR and Northern blotting.
Average Yields of Total RNA Using SV Total RNA Isolation System.
Samples
Mouse Tissues
Liver
Kidney
Spleen
Brain
Muscle
Rat Tissues
Pancreas
Heart
Lung
Bacteria
E. coli
Yeast
S. cerevisiae
Plant
Tomato Leaf
Cell Line
RAW264.7
Maximum
Amount to
Process
Avg. Yield
per Prep
(µg)
Avg. Yield
per mg
Tissue (µg)
A260/A280
30mg
20mg
15mg
60mg
30mg
131
44
79
39
22
4.4
2.2
5.3
0.65
0.73
1.9
1.9
1.9
2.1
2.1
30mg
60mg
60mg
100
16
36
3.5
0.27
0.6
1.9
2.1
2.1
1 x 109 cells
36
N/A
2.0
4 x 107 cells
19
N/A
2.1
30mg
4.6
0.15
2.0
5 x 106 cells
51
N/A
2.1
M
β-actin
5µl
5µl
APC
10µl
Cat.#
Z3101
Z3100
Z3105
15µl
bp
1,500 –
1,000 –
700 –
400 –
RNA isolated from 1ml of human blood using the SV Total RNA Isolation System.
RT-PCR was performed using the indicated volumes of eluted RNA and primers
complementary to human β-actin or human Adenomatous Polyposis Coli (APC) gene
with the Access RT-PCR System(a,b) (Cat.# A1280, A1250).
The vacuum protocol for the SV Total RNA
Isolation System requires use of the
Vacuum Adapters and a vacuum manifold
such as Promega’s Vac-Man ® Laboratory
Vacuum Manifold (20-sample capacity).
Total RNA
• Safety and convenience: Silica membrane based, no phenol
extraction required.
Size
10 preps
50 preps
250 preps
N/A = Not Applicable.
Average yield is that of RNA only. Other commercial systems may appear to give higher yields, but this is
frequently due to DNA carryover.
37
A.
Homogenize samples (tissues,
cultured cells or white blood cells)
in SV RNA Lysis Buffer.
Transfer to fresh tube and add SV RNA
Dilution Buffer; mix and centrifuge.
Transfer to fresh tube;
add 95% Ethanol and mix.
Interslide CV (%)
Spin
Vacuum
Centrifuge for one minute.
Remove lysate by vacuum.
Wash with SV RNA
Wash Solution.
Wash with SV RNA
Wash Solution.
DNase Treatment.
Add SV DNase
Stop Solution.
Centrifuge for one minute.
DNase Treatment. Add
SV DNase Stop Solution.
Wash 2X with SV RNA
Wash Solution.
Wash 2X with
SV RNA Wash Solution.
Elute RNA into
Elution Tube.
Remove Spin Basket and place
in collection tube. Centrifuge.
Place Spin Basket in Elution Tube.
10
9
8
7
6
5
4
3
2
1
0
Exp. 1
Exp. 2
Exp.3
4413TA12_3A
B.
Transfer to Spin Column Assembly.
High degree of experimental reproducibility using RNA purified by the SV Total RNA
Isolation System. Panel A. Subgrids from four representative 4K human cancer
arrays (provided by Corning Incorporated) are shown. Labeled cDNA was generated
with the ChipShot™ components of the Pronto!™ Plus System using 5ug of total RNA
purified using the SV Total RNA System. Panel B. Reproducibility was assessed by
determining the %CV for 4000 features from 3-4 arrays. The inter-slide %CV was
<10% indicating a high degree of reproducibility within and between experiments as a
result of high quality input total RNA.
Total time: 60-70 minutes
Schematic of the SV Total RNA Isolation System protocol.
Promega
1
2
3
4
Competitor
5
6
M
7
8
9
10 11 12
The PCR process is covered by patents issued and applicable in certain countries*. Promega does not
encourage or support the unauthorized or unlicensed use of the PCR process. Use of this product is
recommended for persons that either have a license to perform PCR or are not required to obtain a license.
*In the U.S., effective March 29, 2005, U.S. Pat. Nos. 4,683,195, 4,965,188 and 4,683,202 will expire. In
Europe, effective March 28, 2006, European Pat. Nos. 201,184 and 200,362 will expire.
(b) U.S. Pat. Nos. 4,966,964, 5,019,556 and 5,266,687, Australian Pat. Nos. 616,881 and 641,261 and other
pending and issued patents, which claim vectors encoding a portion of human placental ribonuclease inhibitor,
are exclusively licensed to Promega Corporation.
(a)
38
1906TA09_7A
High-quality RNA from the SV Total RNA
isolation system ensures success in
subsequent experiments such as
microarrays.
Formaldehyde gel electrophoresis of RNA. RNA was isolated from 30mg of liver
tissue following the SV Total RNA Isolation System protocol (lanes 1–6) and a
competitors protocol (lanes 7–12). Ten percent (10µl) of the final prep volume of
100µl for each sample was analyzed by electrophoresis in a 1% MOPS-EDTAformaldehyde agarose gel and visualized by including 0.1µg/µl ethidium bromide in
the sample loading buffer. Promega’s system consistently yielded an average of
100µg. The competitor’s system yielded an average of 70µg with wide variability.
SV 96 Total RNA Isolation System
High-throughput 96-well total RNA purification
The SV 96 Total RNA Isolation System provides:
• Efficiency: Save labor with a walk-away high-throughput 96-well
purification automated method. 96-samples processed is less than
1 hour.
Product
SV 96 Total RNA Isolation System
Size
1 x 96 each
5 x 96 each
Cat.#
Z3500
Z3505
For Laboratory Use.
• A unique design: The novel vacuum manifold eliminates waste
handling, and the novel plate design prevents cross-contamination
during sample processing.
• Confidence in your results: A DNase I treatment step is included
in the protocol to ensure removal of genomic DNA contamination.
There is no detectable DNase carryover into the final RNA
preparation.
• Hassle-free automated methods: Automated methods and
support information available at: www.promega.com/automethods/
3446CA06_1A
• An applications-tested system: Tested to ensure purified RNA
performs optimally in RT-PCR.
Automated isolation of total RNA using the SV 96 Total RNA Isolation System on the
Beckman Coulter Biomek® FX.
0.8
0.7
SV 96
SV
0.6
0.4
0.3
Total RNA
µg RNA
0.5
0.1
0
HeLa
NIH3T3
CHO
3467MB07_1A
0.2
The SV 96 Total RNA Isolation System provides consistent and reliable recovery of
high quality total RNA. Panel A: RNA yields from several cell lines using 1 x 105 cells
is consistently 0.3µg or more.
Total RNA can be purified from 96 samples
at once in less than an hour.
39
MagneSil® Total RNA mini-Isolation System
High-throughput 96-well or 384-well purification of
highly concentrated total RNA
Product
MagneSil® Total RNA
mini-Isolation System
Size
Cat.#
4 plates
Z3351
For Laboratory Use.
The MagneSil Total RNA mini-Isolation System provides:
• A simple, fast method: Elution volumes as low as 15µl give
concentrated RNA. Only 30 minutes to process one 96-well plate
and 50 minutes to process a single 384-well plate on a Beckman
Coulter Biomek® FX laboratory workstation.
Add sample lysate to
MagneSil® RNA PMPs. Mix.
• Confidence in your results: A DNase I treatment step is included
in the protocol to ensure removal of genomic DNA contamination.
• A performance-tested system: RNA rigorously tested in end-point
RT-PCR and quantitative, real-time RT-PCR.
• Efficiency: Save labor with a walk-away high-throughput 96-well
purification automated method. 96-samples processed in only 30
minutes.
• Designed for screening protocols: Total RNA isolation from small
sample sizes (up to 105 cultured cells, 20mg tissue or 20µl whole
blood/well in a 96-well format, and up to 103 cultured cells or 5µl
whole blood/well in a 384-well format).
Capture PMPs.
Discard supernatant.
Wash PMPs.
Incubate PMPs with
DNase 10 minutes.
Add DNase Stop Solution.
• Hassle-free automated methods: Automated methods and
support information available at: www.promega.com/automethods/
Elute RNA.
Overview of the MagneSil Total RNA mini-Isolation System protocol.
40
4316MB11_3A
Capture PMPs.
Discard supernatant.
Wash PMPs.
A.
Yield
GAPDH
40
30
Ct
20
10
1,
00
0
10
,0
00
10
0,
00
0
1E
+0
6
Cell Number
Concentration
60
10
0
silica membrane
10
0
MagneSil® RNA
1
µg
3.0
2.5
2.0
1.5
1.0
0.5
0
40
c-myc
B.
40
20
30
10
0
®
silica membrane
MagneSil RNA
Ct
30
4376MA11_3A
ng/µl
50
20
10
20
10
15
20
25 30
35
40
4378TC
1E
+0
6
10
0,
00
0
10
,0
00
1,
00
0
10
0
45 50
Total RNA
Concentration (ng/µl)
Cell Number
Real-time RT-PCR analysis of RNA purified using the MagneSil Total RNA miniIsolation System. To measure linearity and sensitivity of RNA purification, GAPDH
(abundant mRNA) and c-myc (rare mRNA) were amplified in real-time RT-PCR from
RNA purified from various titrations of HeLa cells. Twenty microliter aliquots of total
RNA isolated from a dilution series of HeLa cells in a 96-well plate were used as
template in 100µl reverse transcription reactions. Panel A: Five microliter aliquots (n=3)
of the RT reaction were used for PCR of a GAPDH target. Panel B: Five microliter
aliquots (n=1) of the RT reaction were used for PCR of a c-myc target. GAPDH signal
and c-myc signal were detected from as little as 10 cells.
30
0
10
10
1
0
Yield of total RNA from HeLa cells using the MagneSil Total RNA mini-Isolation System. Total
RNA was isolated from 1 x 105 HeLa cells/well using either the MagneSil Total RNA mini-Isolation
System or a silica membrane based purification method. Total RNA concentration and yield were
calculated using RiboGreen® RNA Quantitation Reagent (Molecular Probes).
Elution Volume (µl)
0.4
0.2
0.0
10
15
20
25 30
35
40
Elution Volume (µl)
45 50
4377MA11_3A
Yield (µg)
0.6
MagneSil Total RNA mini-Isolation System: titration of elution volume. Total RNA was
isolated from 2 x 104 HeLa cells/well using the MagneSil Total RNA mini-Isolation
System, and eluted in various volumes from 50µl down to 10µl. Yield and concentration
obtained for each elution volume are shown. Total RNA concentration was calculated
using RiboGreen® RNA Quantitation Reagent (Molecular Probes).
41
PolyATtract® Automated System
High-throughput, mRNA purification for sensitive
applications
The PolyATtract Automated System provides:
• Your choice of starting material: Linear recovery of poly(A)+ RNA
from 0.31–20µg of eukaryotic total RNA. Alternatively, work with
0.02–5mg of eukaryotic tissue in lysate or 102–106 of cultured
mammalian cells.
• Strong, selective mRNA binding: A magnetic-based purification
method for mRNA capture that is based on hybridization of
poly(A)+ mRNA to a biotinylated oligo(dT) primer. The bound
mRNA is then captured by streptavidn-coated paramagnetic
particles. Removal of >99% of ribosomal RNA as judged by
quantitative, real-time RT-PCR.
Product
PolyATtract® Automated System
A. 101
100
10–1
10–2
0
10
20
30
40
Cycle
GAPDH Target
Total RNA (Avg. Ct = 23.0)
mRNA From Total RNA (Avg. Ct = 24.9)
• Freedom from well-to-well cross-contamination: No detectable
well-to-well contamination as judged by RT-PCR.
• Hassle-free automated methods: Automated methods and
support information available at www.promega.com/automethods/
Cat.#
Z5671
For Laboratory Use.
• Confidence with expression profiles: Expression profiles remain
the same as samples from total RNA and poly(A)+ RNA.
• Efficiency: Save labor with walk-away automation of mRNA
purification directly from cultured cells, tissue lysates or purified
total RNA.
Size
4 x 96
mRNA From Cells (Avg. Ct = 23.8)
B. 101
100
10–1
0
10
20
30
40
Cycle
1:10 Total RNA
1:100 Total RNA
mRNA From Total RNA
mRNA From Cells
1:1,000 Total RNA
1:10,000 Total RNA
4073TA03_3A
18S rRNA Target
Detection of Ten copies of GAPDH and c-myc using real-time RT-PCR analysis of
RNA purified using the MagneSil Total RNA Mini-Isolation System. To measure
linearity and sensitivity of RNA purification, GAPDH (abundant mRNA) and c-myc (rare
mRNA) were amplified in real-time RT-PCR from RNA purified form various titrations
of HeLa cells. Twenty microliter aliquots of total RNA isolated from a dilution series of
HeLa cells in a 96-well plate were used as template in 100_l reverse transcription
reactions. Panel A: Five microliters aliquots (n=3) of the RT reaction were used for
PCR of a GAPDH target. Panel B: Five microliter aliquots (n=1) of the RT reaction were
used for PCR of a c-myc target.
42
PolyATtract® mRNA Isolation Systems
Scalable, flexible mRNA purification for all
applications
The PolyATtract mRNA System provides:
• Speed: mRNA isolation is completed in 45 minutes or less.
• Strong, selective mRNA binding: A magnetic-based purification
method for mRNA capture that is based on hybridization of
poly(A)+ mRNA to a biotinylated oligo(dT) primer. The bound
mRNA is then captured by streptavidin-coated paramagnetic
particles. Removal of >99% of ribosomal RNA as judged by
quatitative, real-time RT-PCR.
Product
PolyATtract® System 1000
with Magnetic Stand
PolyATtract® System 1000
without Magnetic Stand
PolyATtract® mRNA Isolation
with Magnetic Stand
PolyATtract® mRNA Isolation
(Refill for Z5200)
PolyATtract® mRNA Isolation
with Magnetic Stand
PolyATtract® mRNA Isolation
(Refill for Z5300)
Size
Cat.#
Scalable
Z5420
Scalable
Z5400
3 isolations
Z5200
3 isolations
Z5210
15 isolations
Z5300
15 isolations
Z5310
System II
System I
System III
System IV
For Laboratory Use.
• Your choice of starting material: Configurations available for large
or small amounts of cells or tissues. mRNA isolation from total
RNA in low throughput format (PolyATtract Systems I-IV) or
directly from crude cell or tissue lysates in scaleable format
(PolyATtract System 1000).
• Convenience and ease-of-use: mRNA isolation accomplished
easily, with a simple, magnetic-based procedure.
• Applications tested: Highly pure mRNA suitable for molecular
biology applications such as in vitro translation, cDNA synthesis,
PCR, RNase protection, primer extension and Northern blots.
Average Yields of Poly(A)+ RNA Using PolyATtract System 1000.
Size of
Sample
Yield of
mRNA
A260/A280
µg mRNA/
g sample
Liver
Kidney
Spleen
Pancreas
Lung
NIH3T3
1,000mg
420mg
390mg
400mg
160mg
6 × 107 cells
220µg
138µg
282µg
148µg
16µg
20µg
2.08
2.10
2.15
2.11
2.09
2.12
220
328
72
370
100
200
Total RNA
Sample
(a)U.S.
(b)U.S.
Pat. No. 5,693,784.
Pat. No. 6,218,531, Australian Pat. No. 745185 and other patents pending.
43
Ordering Information
Product
RNAgents® Total RNA Isolation System*
RNAgents® Denaturing Solution*
Product
SV Total RNA Isolation System(a,b)*
SV RNA Red Blood Cell Lysis Solution*
SV RNA Lysis Buffer*
Product
SV 96 Total RNA Isolation System(a)*
SV RNA Lysis Buffer*
SV RNA Wash Solution* (concentrated)
Nuclease-Free Water*
Size
Scalable
120ml
Cat.#
Z5110
Z5651
Size
10 preps
50 preps
250 preps
200ml
50ml
Cat.#
Z3101
Z3100
Z3105
Z3141
Z3051
Size
1 x 96 each
5 x 96 each
50ml
58.8ml
150ml
Cat.#
Z3500
Z3505
Z3051
Z3091
P1195
Hard Ware Devices/Accessories
Product
Vac-Man® Laboratory Vacuum Manifold,
20-sample capacity
Vacuum Adapters
Vac-Man® 96 Vacuum Manifold
Wizard® SV 96 Binding Plates
MagneSil® Magnetic Separation Unit
MagnaBot® 96 Magnetic Separation Device
Heat Transfer Block
MagnaBot® Spacer 1/8 inch
MagnaBot® Spacer
1/4 inch Foam Spacer
PolyATtract® System 1000 Magnetic
Separation Stand
Size
4 plates
Cat.#
Z3351
Product
Size
PolyATtract® Automated System*
4 x 96
Nuclease-Free Water*
150ml
Biotinylated Oligo(dT) Probe (50pmol/µl)*
35µl
Steptavidin MagneSphere Paramagnetic Particles*
25ml
PolyATtract® GTC Extraction Buffer*
120ml
Cat.#
Z5671
P1195
Z5261
Z5482
Z5531
Product
PolyATtract® System 1000 with
Magnetic Stand(c)*
PolyATtract® System 1000 without
Magnetic Stand(c)*
Product
PolyATtract® mRNA Isolation
with Magnetic Stand*
PolyATtract® mRNA Isolation
(Refill for Z5200)
PolyATtract® mRNA Isolation
with Mangetic Stand*
PolyATtract® mRNA Isolation
(Refill for Z5300)
Size
Cat.#
Scalable
Z5420
Scalable
Z5400
Size
Cat.#
3 isolations
Z5200
3 isolations
Z5210
15 isolations
Z5300
15 isolations
Z5310
(a)U.S.
MagneSil is a registered trademark of Promega Corporation.
ChipShot and Pronto! are trademarks of Promega Corporation.
Cy is a registered trademark of Amersham Biosciences Ltd.
Biomek is a registered trademark of Beckman Coulter, Inc.
RiboGreen is a registered trademark of Molecular Probes, Inc.
System I*
System III with
System IV*
Promega Corporation • 2800 Woods Hollow Road • Madison, WI 53711-5399 USA • Telephone 608-274-4330 • Fax 608-277-2601
44
each
each
each
pack
each
each
each
each
each
each
each
A7231
A1331
A2291
A2271
A2278
A2231
V8151
Z3271
V8581
V8381
Z3301
1 each
Z5410
1
20
1
10
100
1
1
1
1
1
1
Pat. No. 6,218,531, Australian Pat. No. 745185 and other patents pending.
Pat. No. 730718 and other patents and patents pending.
(c)U.S. Pat. No. 5,693,784.
System II with
©2004 Promega Corporation. All Rights Reserved.
Prices and specifications subject to change
without prior notice.
Cat.#
Cat.# P1195, Z5261, Z5482, Z5531 Cat.# A2271, A2278, Z3051, Z3091, P1195. For Laboratory Use.
(b)Australian
Product
MagneSil® Total RNA mini-Isolation System(a)*
Size
Printed in USA 09/04
11268-BR-PU
Part #BR106