HCY™ KlenTaq DNA Polymerase

HCY™ KlenTaq DNA Polymerase is a thermally stable recombinant Taq DNA polymerase from the thermophilic bacterium Thermus aquaticus, genetically engineered to have a molecular weight of 62 KD.The enzyme has a deletion of 278 amino acids at the N-terminal end, and compared to the wild-type Taq enzyme, truncated Klentaq enzyme offers higher fidelity and thermal stability. The enzyme retained the DNA polymerase activity of DNA polymerase I and 3’→5′ nucleic acid exonuclease activity, and lacked 5’→3′ nucleic acid exonuclease activity. The product carries an A base at the 3′ end and can be directly cloned with TA vector.

Conventional PCR, high-fidelity PCR, genotyping and primer extension analysis, etc.
HCY™ KlenTaq enzyme can amplify DNA fragments up to 5 kb in length. It is usually suitable for amplification of DNA fragments up to 3 kb in length.
Most PCR products amplified with this product have an “A” base at the 3′ end and can therefore be cloned directly into the T-Vector.

Frequently Asked Questions:

1. No amplified bands

(1) Enzyme is inactivated or not added to the reaction system.DNA polymerase is inactivated by improper storage or transportation.Satisfactory results are often obtained by replacing the enzyme with a new one or with one from another source.
(2) The template contains impurities. In particular, formaldehyde-fixed and paraffin-embedded tissues often contain formic acid, causing DNA depurination and affecting the results of PCR.
(3) Whether the denaturation temperature is accurate: the PCR instrument indicates whether the temperature and the actual temperature is consistent, too high enzyme in the first few cycles of rapid inactivation; too low, the template denaturation is not complete.
(4) The reaction system is contaminated with protease and nuclease, the reaction system should be heated at 95℃ for 5-10 minutes before adding Taq enzyme.
(5) Deterioration and failure of primers. Are the primers synthesized correctly. Are they purified or inactivated due to improper storage conditions.
(6) Primer error. Use BLAST to check primer specificity or redesign primers.
(7) Add a positive control during DNA gel electrophoresis to ensure that it is not a problem with the DNA gel or PCR program.

2. Too little PCR product

(1) Inappropriate annealing temperature. Design a gradient PCR reaction to optimize the annealing temperature with a gradient of 2 degrees.
(2) The amount of DNA template is too small. Increase the amount of DNA template.
(3) Insufficient PCR cycles. Increase the number of reaction cycles.
(4) Insufficient amount of primer. Increase the amount of primer in the system.
(5) Extension time is too short. Set the extension time at 1kb/minute.
(6) Denaturation time is too long. Too long a denaturation time can lead to inactivation of DNA polymerase.
(7) Inhibitors are present in the DNA template. Make sure the DNA template is clean.