2xEs Taq MasterMix

Item Number

E665629

• Storage Temp: 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 does not contain dyes. After the PCR program is completed, an appropriate amount of sample buffer can be added as needed for electrophoresis operation. 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.

E665629	Component	5ml	25ml	Storage
E665629A	2×Es Taq MasterMix	5×1ml	5×5ml	-20℃. Avoid freeze/thaw cycle.
E665629B	ddH₂O	5×1ml	5×5ml	-20℃. Avoid freeze/thaw cycle.

Notes: 2×Es Taq MasterMix contains Es Taq DNA Polymerase, 3mM MgCl2 and 400µM each dNTP.

Quality control:
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.
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	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.