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How to choose a hot-start Taq/DNA polymerase

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How to choose a hot-start Taq/DNA polymerase

Hot-start Taq enzyme is widely used. Compared with ordinary DNA polymerase, hot-start Taq enzyme can effectively avoid some non-specific amplification and the formation of primer dimers, and can effectively improve the success rate of target gene amplification. Especially in the field of genetic testing, hot-start Taq enzyme has been identified as a mandatory standard in the industry, and ordinary DNA polymerase should not be used. As can be seen from the above, hot-start Taq enzymes are widely used. At present, there are many brands of hot-start Taq enzymes on the domestic market, but there are not many hot-start Taq enzymes with high quality. Faced with so many hot-start Taq enzyme products, how should we choose?


1. Select the hot-start Taq enzyme with high amplification efficiency


PCR amplification efficiency is closely related to the performance of Taq enzyme. After a good Taq enzyme reaction system is optimized, the amplification efficiency is above 95%, and the amplification range of the initial template amount is wide. Satisfactory amplification can be obtained when the target gene content is low, and it is not easy to be poisoned when the template amount is high, and the exponential amplification period is long. For the Taq enzyme with poor performance, even if the reaction system has been optimized for many times, the amplification efficiency is still less than 90%, the "S" shape of the amplification curve is not obvious, the slope is small, and the curve is flat. When the amount of template is low, it cannot be amplified, and when the amount of template is high, the amplification effect is not ideal. Therefore, the selection of DNA polymerases with high amplification efficiency is crucial to the success of PCR and qPCR.


2. Select hot-start Taq enzyme with strong enzyme power

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The enzymatic power of the Taq enzyme is related to the amplification efficiency. Generally, the stronger the enzymatic power of the hot-start Taq enzyme, the longer the exponential growth period of PCR amplification, the more typical 'S-shaped' curve, the higher the fluorescence signal value, and the more suitable for multiplex PCR detection. Brand DNA polymerases with weak enzymatic power can generally only support 2-plex reactions. When doing 3-plex reactions, the amplification curve is low, the fluorescence signal value is low, and there is no typical amplification curve, so the results are difficult to judge.


3. Select a hot-start Taq enzyme with high sensitivity


Generally speaking, DNA polymerase has high amplification efficiency and high sensitivity, but there are also inconsistencies. If the target gene abundance of the sample to be amplified is low, it is recommended to test the amplification sensitivity of Taq enzyme. The most common detection method is to carry out 10-fold or 5-fold gradient dilution of the target gene plasmid fragment, carry out PCR detection at the lower dilution, and select the hot-start Taq enzyme with higher detection sensitivity.

It can be seen from the above that researchers need to choose according to their own experimental requirements and funding conditions. It is best to do a gradient dilution amplification experiment to detect the amplification efficiency and sensitivity of the hot-start Taq enzyme.


An example of Foregenes Taq DNA Polymerase:

Foreasy HS Taq DNA Polymerase


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Foreasy HS Taq DNA Polymerase is a DNA polymerase expressed in Escherichia coli engineering bacteria by gene recombination technology. The enzyme is combined with a unique reaction buffffer, which makes the product highly resistant and compatible, and can directly use the sample lysate (Foregene Lysis system) as a template for detection reactions.

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Qualitative PCR and quantitative PCR detection of purifified templates and non-purifified templates.

Quality Control

1. No exogenous nuclease activity detected

2.PCR method to detect residual genomic DNA without host

3.It can effffectively amplify single-copy genes in the human Genome

4.Store at room temperature for a week,noobvious activity changes


Pub Time : 2022-08-05 11:41:16 >> News list
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