Store at -20°C,Avoid repeated freezing and thawing
Shipped In
Ice chest + Ice pads
Grade
EnzymoPure™
Product Description
Es Taq DNA Polymerase is an optimized mixed enzyme of Taq and Pfu DNA Polymerase, with 5 '→ 3' DNA polymerase activity, 5 '→ 3' exonuclease activity, and 3 '→ 5' exonuclease activity. Compared with Taq DNA Polymerase, Es Taq DNA Polymerase has excellent performance of high amplification efficiency and low mismatch rate, and can efficiently amplify DNA fragments. Most of the PCR products amplified with this product contain an "A" base at the 3 'end, which can be directly used for T/A cloning. This product is suitable for conventional PCR reactions and gene cloning reactions that require high fidelity.
E665597
Component
500 U
Storage
E665597A
Es Taq DNA Polymerase, 5 U/μL
100 μL
-20℃. Avoid freeze/thaw cycle.
E665597B
10×PCR Buffer
1.8 mL
-20℃. Avoid freeze/thaw cycle.
Activity definition: Using activated salmon sperm DNA as a template/primer, the amount of enzyme required to incorporate 10 nmol of deoxyribonucleotide into acidic insoluble substances is defined as 1 active unit (U) at 74 ℃ for 30 minutes. Quality control: After multiple column purifications, SDS-PAGE detected a purity of over 99%; No exogenous nuclease activity detected; PCR method for detecting residual DNA without host; Can effectively amplify single copy genes in the human genome; Store at room temperature for one month without significant changes in activity.
1. PCR reaction system
Reagent
50 μlReaction system
Final concentration
10×PCR Buffer
5 μL
1×
dNTP Mix，10 mM each
1 μL
200 μM each
Forward Primer，10 μM
2 μL
0.4 μM
Reverse Primer，10 µM
2 μl
0.4 μM
Template DNA
<0.5 μg
<0.5 μg/50 μl
Es Taq DNA Polymerase，5 U/μl
0.25-0.5 μl
1.25-2.5U/50 μl
ddH2O
up to 50 μL
/
Attention: The primer concentration should be between 0.1 and 1.0 as the final concentration μ M serves as a reference for setting the range. In the case of low amplification efficiency, the concentration of primers can be increased; When non-specific reactions occur, the primer concentration can be reduced to optimize the reaction system.
2. PCR reaction conditions
Step
Temperature
Time
/
Pre denaturation
94℃
2 min
/
Denaturation
94℃
30 s
25-35 cycles
Anneal
55-65℃
30 s
25-35 cycles
Extend
72℃
30 s
25-35 cycles
Finally extended
72℃
2 min
/
Attention: 1) In general experiments, if the annealing temperature is 5 ℃ lower than the melting temperature Tm of the amplification primer, and the ideal amplification efficiency cannot be achieved, the annealing temperature should be appropriately reduced; When non-specific reactions occur, increase the annealing temperature to optimize the reaction conditions. 2) The extension time should be set according to the size of the amplified fragment. The amplification efficiency of Es Taq DNA Polymerase in this product is 2 kb/min. 3) The number of cycles can be set based on the downstream application of the amplification product. If the number of cycles is too small, the amplification amount is insufficient; If there are too many cycles, the probability of mismatches will increase, and non-specific backgrounds will be severe. So, while ensuring product yield, the number of cycles should be minimized as much as possible.
