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method for improving PCR amplification efficiency and amplification length of DNA high fidelity polymerase

A high-fidelity polymerase and ultra-long technology, which is applied in the field of improving the amplification efficiency and amplification length of DNA high-fidelity polymerase PCR, and can solve problems such as amplification and complex templates

Inactive Publication Date: 2019-06-18
中禾生物种业集团有限公司
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  • Application Information

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Problems solved by technology

[0006] The purpose is to solve the process of using high-fidelity DNA polymerase to amplify, how to further increase the activity of high-fidelity DNA polymerase, and solve the problems faced by high-fidelity DNA polymerase in amplifying complex templates and long fragments

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  • method for improving PCR amplification efficiency and amplification length of DNA high fidelity polymerase
  • method for improving PCR amplification efficiency and amplification length of DNA high fidelity polymerase

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Embodiment Construction

[0009] Explored high-fidelity DNA polymerase amplification conditions: In the amplification system, take the 20ul amplification system as an example, add 0.5ul Triton X-100 into the PCR reaction solution, and add a small amount of 0.5ul Triton X-100 Hypromellose Magnesium Chloride Buffer. The formula of the hydroxypropylmethylcellulose magnesium chloride buffer solution is 1L of deionized water, containing 15 grams of hydroxypropylmethylcellulose and 10 grams of magnesium chloride. Before the PCR experiment, add hypromellose magnesium chloride buffer and PCR buffer on ice to pre-mix, and then pre-mix with PCR reaction enzyme. In the final PCR, the buffer, DNA template, primers, etc. are mixed. Other amplification conditions are set according to the template. The primer design and the reaction conditions of the high-fidelity enzyme are consistent with the amplification conditions of the normal high-fidelity DNA polymerase.

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Abstract

The invention provides a method for improving the PCR amplification efficiency of a DNA high fidelity polymerase. According to the method, by adding Triton X-100, hydroxypropyl methyl cellulose and amagnesium chloride buffer solution to a PCR high fidelity enzyme reaction system, the amplification efficiency of the high fidelity DNA polymerase is improved; and the amplification length, amplification speed and amplification efficiency of ordinary high fidelity DNA polymerase are improved, and the advantages of simpleness and convenience, rapidity and advantageous use are achieved.

Description

technical field [0001] This method involves the amplification of a high-fidelity DNA polymerase. Even if the reaction buffer and the sample loading steps of the high-fidelity DNA polymerase are improved, the maximum DNA amplification efficiency of the high-fidelity DNA polymerase can be achieved. Background technique [0002] PCR is a very commonly used technique in modern molecular biology techniques, and many technical methods and improved application techniques have been derived from this technique. PCR is a method of using polymerase to amplify gene DNA fragments. In the process of continuous heating, annealing and cooling, artificially designed targeting primers can bind to the target DNA with the help of polymerase to achieve targeting only amplifies the specific fragment of interest. Using this technology, the concentration of the target gene for research can be greatly accelerated, and the processing and manipulation of downstream DNA fragments is convenient. [00...

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Application Information

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IPC IPC(8): C12Q1/686
Inventor 不公告发明人
Owner 中禾生物种业集团有限公司