2xTaq MasterMix（Dye）

Item Number

T665590

• Storage Temp: Store at -20°C,Avoid repeated freezing and thawing
• Shipped In: Ice chest + Ice pads

Product Description

Taq MasterMix is a premixed system composed of Taq DNA Polymerase, Mg2+, dNTPs, as well as PCR stabilizers and enhancers, with a concentration of 2 ×. The pre prepared PCR mixture makes the operation simpler and faster, and can minimize human error and contamination to the greatest extent possible. The original MasterMix formula results in high yield, strong repeatability, and good stability of amplified products. This product has been added with a dye (blue), and can be directly subjected to electrophoresis detection after the reaction is completed. The amplified PCR product has an "A" base attached to its 3 'end, making it suitable for direct use in T/A cloning. Mainly suitable for PCR amplification of DNA, DNA sequencing and other experiments.

T665590	Component	5 mL	25 mL	Storage
T665590A	2×Taq MasterMix (Dye)	5×1 mL	5×5 mL	-20℃. Avoid freeze/thaw cycle.
T665590B	ddH₂O	5×1 mL	5×5 mL	-20℃. Avoid freeze/thaw cycle.

2×Taq MasterMix contains Taq DNA Polymerase, 3 mM Mg Cl₂ and 400 µM 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×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	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 ° C 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 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.