Taq-red DNA Polymerase
User’s Instruction
Description
Taq-red DNA Polymerase is an unique blend of U-Taq DNA Polymerase with
an inert red dye. It offers the same performance as U-Taq DNA Polymerase.
Several benefits are as follows:
This red dye enables quick and convenient visual confirmation of enzyme
addition and reaction mixing.
1. After PCR amplification, samples can be removed from the reaction and
loaded directly onto agarose gel without the addition of loading buffer or
tracking dye. The red dye, acting as a tracking dye, migrates between
bromophenol blue and xylenecyanol at about the same rate as 400~500
bp fragment.
2. The concentration of Taq-red is only 1unit/µl, so the enzyme can be
transferred and added more accurately.
3. The enzyme generates PCR products with 3'-dA overhangs, suitable for
T-A cloning.
4. 6kb Lambda DNA and 2.1 kb Human genomic DNA can be amplified very
well at our laboratory.
Contents
500 units
1. Taq-red DNA Polymerase (1 unit/μl)
100 μl
2. 10×PCR reaction buffer
1.5 ml
Taq-red DNA Polymerase Storage Buffer: Taq DNA Polymerase is supplied
in 50mM Tris-HCl (pH8.0), 100mM NaCl, 0.1mM EDTA, 0.5mM DTT,
1%TritonX-100, and 50%Glycerol.
10×PCR Reaction Buffer: 500mM KCl, 100mM Tris-HCl (pH8.0), 20mM
MgCl2.
Note: Optimized 10×PCR reaction buffer is available with or without MgCl2, A
separate tube of 20mM MgCl2 is included with the 10×PCR reaction buffer not
containing MgCl2.
Protocol
The following is an example of PCR reaction, only for reference.
1. In a 0.2 ml or 0.5 ml thin wall tube, assemble in order:
20 μl reaction
50 μl reaction
10×PCR reaction buffer
2μl
5μl
Forward primer
10-20 pmol
20-50 pmol
Reverse primer
10-20 pmol
20-50pmol
Template DNA
5-50 ng
10-100 ng
dNTPs (10mM each)
0.5μl
1 μl
Taq-red DNA Polymerase
1-1.5 unit
2-3units
ddH2O
X μl
X μl
Total
20μl
50μl
2. Mix gently by pipetting, close the tube and centrifuge for a few seconds.
3. Add mineral oil to each tube (This step is unnecessary when using a
thermal cycler with top heating.).
4. Perform PCR cycles according to the PCR condition.
(Annealing temperature and time need to be optimized for each primer/
template combination.)
94℃
2.5 minutes
94℃
45 seconds
50~65℃
1 minute
72℃
1-3 minutes
72℃
5-10 minutes
25-35 cycles
Storage Conditions
Store the enzyme at -20℃. The enzyme is stable for more than one year under
suggested storage condition. Avoid multiple freeze-thaw cycles and exposure
to frequent temperature changes.