Store at -20°C,Avoid repeated freezing and thawing
Shipped In
Ice chest + Ice pads
Product Description
This product is a premixed system composed of Es Taq DNA Polymerase, Mg2+, dNTPs, PCR stabilizers and enhancers, with a concentration of 2 ×. Es Taq DNA Polymerase has excellent performance of high amplification efficiency and low mismatch rate. The unique MasterMix formula makes the entire reaction system very stable, with over 98% of PCR amplification successful at once. At the same time, complex templates can also be effectively amplified, and human error and contamination can be minimized to the greatest extent. This product has been added with a dye (blue), and can be directly subjected to electrophoresis detection after the reaction is completed. Most PCR products obtained from amplification have an "A" base attached to the 3 'end, making them suitable for direct use in T/A cloning. Mainly suitable for routine PCR reactions and gene cloning experiments that require high fidelity.
After testing, there was no exogenous nuclease activity; PCR method for detecting residual DNA without host; Can effectively amplify single copy genes from multiple genomes.
E665625
Component
1 mL
5 mL
25 mL
40 mL
Storage
E665625A
2×Es Taq MasterMix (Dye)
1 mL
5×1 mL
5×5 mL
40×1 mL
-20℃. Avoid freeze/thaw cycle.
E665625B
ddH2O
1 mL
5×1 mL
5×5 mL
40×1 mL
-20℃. Avoid freeze/thaw cycle.
Notes: 2×Es Taq MasterMix contains Es Taq DNA Polymerase, 3mM MgCl2 and 400µM each dNTP.
Usage: The following is an example of a PCR reaction system and reaction conditions for amplifying a 1 kb fragment using human genomic DNA as a template. In practical operation, corresponding improvements and optimizations should be made based on the template, primer structure, and target fragment size. 1. PCR reaction system
Reagent
50 μlReaction system
Final concentration
2×Es Taq MasterMix（Dye）
25 μl
1×
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
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
95℃
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, and the amplification efficiency of Es Taq DNA Polymerase 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.
