Academic Treasure Land: Applicable Scenarios of Different Types of Taq Enzymes

It has 5 '-3' polymerase activity and 5 '-3' exonuclease activity, but does not have 3 '-5' exonuclease activity.

It is mainly applied in simple gene fragment amplification, for example: 1) Obtaining specific gene fragments from known templates for subsequent preliminary research such as cloning and expression analysis; 2) To study the expression of a specific gene in a certain plant, the gene fragment was amplified by PCR. The common Taq enzyme can rapidly and massively replicate the target DNA, providing sufficient materials for subsequent experiments. 3) In genotype identification experiments, by amplifying specific gene loci, an individual's genotype is determined based on the size and sequence characteristics of the amplification products.

The mismatch rate of common Taq enzymes is relatively high, approximately 10⁻⁵ bases per cycle number⁴This means that some incorrect bases may be introduced during the amplification process, so it is not very suitable for experiments with extremely high fidelity requirements.

The emergence of high-fidelity enzymes is precisely to address the high mismatch rate of common Taq enzymes. Its mismatch rate can be reduced to the order of 10⁻6, which is attributed to its 3 '-5' exonucidase activity. It can correct the wrongly incorporated bases during DNA synthesis, just like a strict "calibrator", ensuring the high accuracy of the amplified DNA sequence.

The most distinctive feature of the hot-start Taq enzyme is that it has no activity at room temperature and only takes effect after being activated at high temperatures. This characteristic greatly enhances amplification in PCR experimentsSpecificity and stability under fully premixed conditions with probing。

Take the M2071 fully premixed DNA system modified by Baorui Antibody as an example. It can support stability at 37℃ for 14 days with the introduction probe. Stable at 2-8℃ and 12M.

In fields such as genomic mapping, sequencing, and molecular genetics research, researchers often need to amplify extremely long DNA fragments. Due to its own characteristics, ordinary Taq enzymes find it difficult to complete this arduous task, while ultra-long fragment amplification Taq enzymes are specifically designed for this purpose. The ready-to-use 2× high-fidelity PCR premix has a fidelity approximately 96 times that of Taq enzyme. Unique extension factors and specific promoting factors were added to the reagent, which significantly enhanced its long-fragment amplification ability, amplification specificity and amplification yield.

Simple templates such as λDNA and plasmids can effectively amplify fragments up to 30kb. Using complex templates such as genomic DNA, fragments up to 20 kb in length can be amplified. In addition, it has good resistance to PCR inhibitors and can be used for direct PCR of bacteria, fungi, plant tissues, whole blood samples or crude nucleic acids.

This product is also available in a version containing electrophoresis indicators, allowing for direct spotting and electrophoresis after the PCR reaction is completed. Experiments such as gene cluster cloning and ultra-long fragment assembly can be easily carried out.

The rapid amplification enzyme is obtained through high-throughput evolutionary screening, and its amplification rate can reach more than five times that of Taq enzyme.