GB-Amp™ HotStart Taq DNA polymerase Gene Bio Systems

Description

GB-Amp™ HotStart Taq DNA polymerase is a hot-start polymerase with chemical modification, which brings higher specificity by reducing non-specific products as the enzyme activity is temperature-dependent and is inhibited at room temperature. The amplification length and speed can reach to 5 kb (simple template) and 0.5 kb/min separately. Hotstart Taq has 5’-3’ polymerase activity, but no 3’-5’ exonuclease activity. The products of Taq plus have overhanged dA at 3’-end.

Components: Hotstart Taq DNA polymerase (5U/ul); 10 Hotstart Buffer (including Mg), 6 x gel loading buffer

 

Stable for 2 years at -20°C

GB-Amp™ HotStart Taq DNA polymerase has zero animal source pollution by being produced with advanced chemical modification. And it is much more stable than antibody-modified hot-start polymerase. Its efficiency is higher than most chemical-modified polymerase and the initial-denaturation time can be reduced to 3 minutes.

 

Unit Definition

One enzyme unit (U) refers to the amount of enzyme needed for intaking 10 nmol nucleotides when using activated salmon sperm DNA as template/primer, at 72 ℃, in 30 minutes

 

Quality control

The absence of endonuclease or exonuclease is confirmed by appropriate quality tests. PCR detects no host residual DNA, and it can effectively amplify the single-copy gene of human genome. There is no significant change about the amplification activity after one week at room temperature.

Storage Buffer

200 mM Tris-HCl, 1 mM DTT, 0.1 mM EDTA, 100 mM KCl, 0.5% Tween 20, 50% Glycerol, 0.5% Triton X -100

 

Notes

1 This product adopts improved chemical modification technology, so it relies on temperature to activate the polymerase activity, which can effectively inhibit non-specific bindings, and the reaction system can be configured at room temperature.

2 This Hotstart Taq DNA polymerase has the deoxidizing nucleotide transfer activity, so at the 3’-end of the PCR product is usually added to an extra deoxygenated adenosine nucleoside.

 

Endodeoxyribonuclease Assay

No detectable conversion of covalently closed circular DNA to a nicked DNA was observed after incubation of 10U GB-Amp™ HotStart Taq DNA polymerase with 1μg pBR322 DNA for 4 hours at 37°C and 70°C.

 

Exodeoxyribonuclease Assay

No detectable degradation of lambda DNA-HindIII fragments was observed after incubation of 10U GB-Amp™ HotStart Taq DNA polymerase with 1μg digested DNA for 4 hours at 37°C and 70°C.

 

Ribonuclease Assay

0% of the total radioactivity was released into trichloroacetic acid-soluble fraction after incubation of 10U GB-Amp™ HotStart Taq DNA polymerase with 1μg E.coli [3H]-RNA (40000cpm/μg) for 4 hours at 37°C and 70°C.

 

Q&A

Why the specificity of GB-Amp™ HotStart Taq DNA polymerasee is higher than ordinary Taq DNA polymerase?

The Hotstart Taq DNA polymerase is a chemically modified Taq DNA polymerase, which activity is completely inhibited at room temperature and it is activated at high temperature in a very short time. Low temperature inhibition & high temperature activation keep the non-specific reactions, i.e., primer dimer formation and off-target amplification low, and thus the specificity high.

 

Are the products of Hotstart Taq DNA polymerase compatible to TA cloning?

Yes. Because the products have 3’-dA protruding ends.