PCR Related Products
Current Products

Thermostable DNA polymerases
– Pfu
– PfuTurbo
– TaqPlus Precision
– TaqPlus Long
– Taq

TaqExtender
Perfect Match
 Opti-Prime

Components of PCR
Thermostable DNA polymerase
 Reaction Buffer
 Primers
 Template DNA
 dNTPs
 Water

– Cycling Conditions
PCR Qualities
Yield
 Fidelity
 Specificity
 Length

Thermostable DNA
polymerases
Pfu
 PfuTurbo
 TaqPlus Precision
 TaqPlus Long
 Taq

error rate
(x10 -6 )
PfuTurbo DNA
polymerase
1.3
+ 0.2
extension
time
required
(min/kb
target)
Recommended
Length
PCR Target
(kb)
vector
targets
genomic
targets
1
0-10
0-6
2
0-15
0-10
Pfu DNA
polymerase
1.3
+ 0.2
2
0-10
0-4
TaqPlus
Precision
system
3.9
+ 1.3
1
0-15
0-10
TaqPlus Long
system
7.6 + 1.2
0.5-1
0-35
0-18.5
Taq DNA
polymerase
8.0 + 3.9
1
0-10
0-5
Functional Domains of Pfu DNA
Polymerase
5’-3’
exo
1
nonfunctional
DNA-dependent DNA
polymerase
3’-5’ exo
126
329
proofreading
activity
775
DNA synthesis
Fidelity Assay

PCR
– not single round extension
Mutations are detected in the lacI gene
 Color detection from Big Blue system
 Results have been published in peer
reviewed journals

Characteristics of Pfu
High Fidelity
 Native Hot Start activity
 High Thermostability
 Slow polymerization rate
 Available as native and cloned enzyme

Accuracy of Polymerases in
PCR
800000
700000
600000
500000
400000
300000
200000
Tma
Tbr
Tfl
Taq
Hot Tub
Vent
Deep Vent
0
Pfu
100000
Extension Time
Pfu requires 2 min/kb
Minutes of extension time/kb of PCR target
M
Taq
Pfu
0.25 0.5
1
1.5
2
2.5
3
3.5
4
Activity vs Temperature
Pfu and Taq
percent
activity
25
35
50
60
75
Incubation Temperature (degrees C)
95
Pfu: Hot Start Every Cycle
M
T
TS
ATG
TMg
Pfu
Polymerase Thermostability
DNA Polymerase
Polymerization Opt
Half Life 95oC
Pfu
70-80oC
19 hours
Taq
70-80oC
1 hour
Characteristics of PfuTurbo
Improved version of Pfu
 Same high fidelity as Pfu
 Higher PCR product yield than Pfu alone
 Can use 1 min/kb extension times
 Works well with low amounts of DNA
template

PCR Enhancing Factor (PEF)
Isolated from Pyrococcus
 Lacks DNA polymerase and exonuclease
activities
 Information on identity/mechanism of
action is proprietary

PfuTurbo vs Other
Polymerases
M PT P
V DV Pw K T M
PT P
V DV Pw K
T
M
5.2 kb
0.9 kb
PfuTurbo vs Pfu
M T P T P T P T P T P
8 kb
6.1 kb
5.2 kb
1.9 kb
0.9 kb
Characteristics of
TaqPlus Precision
Blend of Taq and Pfu
 High Yield
 High Fidelity
 Length
 PCR Buffer

– optimized for both yield and fidelity
180000
160000
140000
120000
100000
80000
60000
40000
20000
0
GeneAm p XL
Advantage Tth
ExTaq
TaqPlus Long
Advantage
KlenTaq
LA PCR
Elongase
Expand Long
Expand High
Fidelity
TaqPlus
Precision
Accuracy of Polymerase Blends in
PCR
PCR with TaqPlus Precision vs Taq
Taq
M
2 5.2
TaqPlus Precision
5.4 10
15
M
2
5.2 5.4 10
15 kb
Characteristics of TaqPlus
Long
Blend of Taq and Pfu
 Long targets
 Fidelity similar to Taq
 Two optimal buffers supplied

– high salt buffer helps prevent smearing in
long target amplification
– low salt buffer
PCR with Taq vs TaqPlus Long
Qualities of Taq
Taq2000 is the recombinant Taq
 Native Taq is also available
 Taq is trusted by customers
 Taq has been well studied

TaqExtender
The Pfu component of TaqPlus Long
 Can be added to any Taq to make a
polymerase blend

Perfect Match PCR Enhancer
Taq2000
M
-
+
native Taq
-
+
Opti-Prime PCR Optimization Kit

12 buffers
– matrix of pH, Mg, KCl

Adjuncts
– DMSO, formamide, BSA, glycerol,
ammonium sulfate, Perfect Match

Recommended for use with Taq
Related PCR products

RoboCycler
– gradient
Nucleotides and primers
 StrataPrep PCR purification kit
 Cloning Vectors

– PCR-Script Kits
– Affinity protein expression kit
QuikChange site-directed mutagenesis
kit
 Cyclist DNA sequencing kits
