This product is a mixture of Taq enzyme, dNTPs, MgCl2 and reaction buffer required for PCR reaction. The 2×Taq Master Mix is optimized for conventional PCR amplification reaction, and the amplification length is up to 8KB, which can efficiently amplify the fragment length of 4KB and below. When using, the template and primer can be added and diluted to 1 times the concentration, which greatly simplifies the operation process and reduces the pollution in the PCR operation process. 2 × Taq Master Mix is available in regular/quick sample format. After testing, the addition of dye does not affect the PCR reaction, after the PCR reaction can be directly electrophoresis, saving time. The PCR product 3´ end amplified with this reagent has A prominent "A" base, which can be directly cloned into T vector.

 

■ Product features:

 

High efficiency: using λDNA as template, the amplification length can be up to 8KB, and the amplification can be no more than 4KB

 

Sensitivity: specific gene fragments can be amplified from the 0.05ng human genomic DNA template.

 

Stability: repeated freezing and thawing dozens of times, placed at 4℃ for 30 days, room temperature after a week, the amplification performance is not affected.

 

Fast: all the reagents necessary for PCR reaction are collected in one tube, and the reaction system can be prepared in a few minutes.

 

Convenience: PCR reaction can be directly electrophoresis (including dye form).

 

■ Product use:

 

1) Conventional PCR identification;

 

2) Small fragment target gene cloning;

 

3) Add A to the flat-end PCR product.

 

S save:

 

-20 ℃, shelf life of 2 years

 

■ Product packaging (A packaging) :

 

Product component volume

 

2 x Taq Master Mix 1 ml x 5

 

PCR Mix 50 mu l

 

*PE012-02 series include upper sample dye

 

 

 

■ Quality assurance:

 

No exogenous nuclease residue was detected and no E. coli DNA residue was detected by qPCR, which can effectively amplify single copy gene in human genome.

 

 

 

 

 

 

 

 

 

 

 

■ Operation method:

 

Common reaction system (50μ L) :

 

2 × Taq Master Mix* 25μ L

 

Upstream primer 0.2-1.0μM (final concentration)

 

Downstream primer 0.2-1.0μM (final concentration)

 

The template

 

 

 

Taq 1-50ng (plasmid)

 

10ng-1μg (genome)

 

0.25 mu (1.25 l U)

 

DdH2O to 50 mu l

 

*Mg2+ final concentration is 2mM

 

Common PCR cycles:

 

When the amplified fragment < 3K;

 

94℃, 1 minute 30 seconds

 

94 ℃, 20 seconds

 

30 cycles: 50 ~ 60℃, 20 seconds

 

72 ℃, 1 KB / 60 seconds

 

72 ℃ for 5 minutes

 

4 ℃ heat preservation

 

 

 

When the amplified fragment ≥3K (recommended primer length ≥30bp);

 

94℃, 5 minutes

 

30 cycles: 94℃, 5 seconds

 

68 ℃, 1 KB / 60 seconds

 

72 ℃ for 5 minutes

 

4 ℃ heat preservation

 

 

 

Matters needing attention

 

1) It needs to be completely dissolved before use to prevent uneven ion concentration.

 

2) The pre-denaturation time ≥5min in PCR was more conducive to wall breaking.

 

3) Appropriate number of cycles should be selected according to the purpose of the experiment, too few cycles will cause insufficient expansion increment; When the number of cycles is too high, the increment of enlargement increases, but the mutation rate also increases, resulting in nonspecific amplification.

 

4) Set the appropriate annealing temperature according to the PRIMER Tm value. If the annealing temperature is too low, non-characteristic amplification will be caused. Too high may not amplify.