Preparation method?of hot start DNA polymerase, prepared polymerase?and application

A polymerase and hot start technology, applied in the biological field, can solve problems such as product contamination, easy DNA damage, incomplete sealing, etc., and achieve the effect of improving amplification specificity and avoiding non-specific amplification

Active Publication Date: 2014-05-28
WUHAN GENECREATE BIOLOGICAL ENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the above technical problems, the present invention provides a method for preparing a hot-start DNA polymerase, the prepared polymerase and its application,...

Method used

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  • Preparation method?of hot start DNA polymerase, prepared polymerase?and application
  • Preparation method?of hot start DNA polymerase, prepared polymerase?and application
  • Preparation method?of hot start DNA polymerase, prepared polymerase?and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Preparation of single-strand DNA-binding protein (SSB) and expression using pET28a(+)-ssb recombinant plasmid (Wang Jianping, Zou Bingjie, Chen Zhiyao, Ma Yinjiao et al. Characterization of recombinant Escherichia coli single-strand binding protein and its improvement in the accuracy of pyrosequencing Sexual application. Acta Biological Engineering. 27 (10): 1513-1520) or purchased through commercial channels. In addition, the preparation of Taq DNA polymerase monoclonal antibody and the preparation of Taq DNA polymerase can be prepared by referring to existing literature reports, or can be obtained through commercial channels (such as purchased through Shanghai Jingjing Molecular Biotechnology Co., Ltd.).

[0020] Measure the corresponding single-stranded DNA binding protein, Taq DNA polymerase monoclonal antibody, and DNA polymerase according to the components shown in the table below, mix them evenly and store them for later use to obtain the hot-start DNA polymerase....

Embodiment 2

[0023] The methods for obtaining single-stranded DNA binding protein, Taq DNA polymerase monoclonal antibody and DNA polymerase are the same as in Example 1.

[0024] Measure the corresponding single-stranded DNA binding protein, Taq DNA polymerase monoclonal antibody, and DNA polymerase according to the components shown in the table below, mix them evenly and store them for later use to obtain the hot-start DNA polymerase.

[0025] Taq DNA polymerase

Embodiment 3

[0027] The methods for obtaining single-stranded DNA binding protein, Taq DNA polymerase monoclonal antibody and DNA polymerase are the same as in Example 1.

[0028] Measure the corresponding single-stranded DNA binding protein, Taq DNA polymerase monoclonal antibody, and DNA polymerase according to the components shown in the table below, mix them evenly and store them for later use to obtain the hot-start DNA polymerase.

[0029] Taq DNA polymerase

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Abstract

The invention relates to a preparation method?of a hot start DNA polymerase,?comprising the following steps:?obtaining a?single stranded DNA binding protein, a Taq DNA polymerasemonoclonal antibody?and?a Taq DNA polymerase; and mixing the?single stranded DNA binding protein, the Taq DNA polymerase?monoclonal antibody?and the Taq DNA polymerase in accordance with?aproportion. The contents of the?single stranded DNA binding protein, the Taq DNA polymerasemonoclonal antibody?and the Taq DNA polymerase are all 20-40%?respectively. The invention also relates to the hot start DNA polymerase prepared by the above method and an application thereof. The hot start DNA polymerase provided by the invention can also be used to solve problems which appear easily in hot start PCR technology such as DNA damage, product contamination?and bad effect of hot start caused by incomplete sealing.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preparing a hot-start DNA polymerase, the prepared polymerase and an application thereof. Background technique [0002] In the application of PCR technology, there are problems such as the appearance of non-specific amplification products, the formation of primer-dimers or the absence of amplification bands on gel electrophoresis. [0003] At present, the above technical problems are usually solved by hot-start PCR technology. In hot-start PCR technology, a specific hot-start DNA polymerase is used to help realize the process of hot-start PCR, but in the conventional hot-start PCR reaction process, it is necessary to modify the reaction system. The components in it are blocked to ensure the smooth progress of the hot-start PCR reaction process. Commonly used hot-start PCR blocking methods include chemical blocking, physical isolation, and DNA-binding protein inhibition....

Claims

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Application Information

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IPC IPC(8): C12N9/12C12N15/10
CPCC12Q1/6848C12Q2521/101C12Q2522/101C12Q2527/127
Inventor 华权高易汪雪马锋沈鹤霄
Owner WUHAN GENECREATE BIOLOGICAL ENG CO LTD
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