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Method for improving reaction efficiency of nucleic acid in vitro amplification

A nucleic acid amplification reaction and in vitro amplification technology, which is applied in the field of improving the efficiency of nucleic acid in vitro amplification reactions, can solve the problems of false positives of multiple primers, reduced sensitivity, high sensitivity, and the experimental environment is easily contaminated by aerosols, and achieves the effect. Excellent, high-sensitivity, high-sensitivity results

Pending Publication Date: 2019-11-08
SHANGHAI NAT ENG RES CENT FORNANOTECH
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0004] In view of the problems encountered in the existing nucleic acid in vitro amplification technology due to the high sensitivity, the experimental environment is easily polluted by aerosols, and non-specific binding occurs between multiple primers, it is easy to produce false positives and reduce the sensitivity. The purpose of the present invention is to: provide A method for improving the reaction efficiency of nucleic acid amplification in vitro

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  • Method for improving reaction efficiency of nucleic acid in vitro amplification
  • Method for improving reaction efficiency of nucleic acid in vitro amplification
  • Method for improving reaction efficiency of nucleic acid in vitro amplification

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Embodiment 1

[0021] A method for improving the reaction efficiency of nucleic acid in vitro amplification, by adding carbon quantum dot materials to the nucleic acid in vitro amplification reaction system to realize the optimization of nucleic acid in vitro amplification, according to the following steps:

[0022] Optimization of carbon quantum dots to PCR amplification reaction system, wherein:

[0023] (1) Prepare a nucleic acid amplification reaction system (except carbon quantum dots), the composition of which is as follows:

[0024]

[0025] Among them, primers P1 and P2 are Staphylococcus aureus-specific nucleic acid amplification primers, and the template is Staphylococcus aureus DNA (from China General Microorganism Culture Collection Center CGMCC 1.2465, and the strains were cultivated using bacteria from Beijing Tiangen Bioengineering Company Nucleic acid extraction kit to extract genomic DNA), and the amount of template was serially diluted.

[0026] (2) Add treated optimize...

Embodiment 2

[0030] A method for improving the reaction efficiency of nucleic acid in vitro amplification, by adding carbon quantum dot materials to the nucleic acid in vitro amplification reaction system to realize the optimization of nucleic acid in vitro amplification, according to the following steps:

[0031] Optimization of carbon quantum dots for ring-mediated isothermal amplification reaction system of carp spring viremia virus, in which,

[0032] (1) Prepare a nucleic acid reaction system, the composition of which is as follows, where primers FIP, BIP, F3 and B3 are specific nucleic acid amplification primers for carp spring viremia virus (the loop primer LF is not added to the system), and the template is carp spring viremia virus virus cDNA. The sample volume of carbon quantum dots is 2.0 µL (final concentration is 20 μg / L):

[0033]

[0034] (2) Add treated optimized material carbon quantum dots to the above system, add 2.0 µL of carbon quantum dot solution to each 25 µL sy...

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Abstract

The invention discloses a method for improving nucleic acid (nucleic acid) amplification reaction efficiency, and belongs to the technical field of biology. According to the method provided by the invention, carbon quantum dots are added to a reaction system, and nucleic acid in vitro amplification is performed. The method can effectively improve the sensitivity of a nucleic acid in vitro amplification reaction, can restrain the appearance of a false positive result, and can reduce non-specific amplification, so that the detection efficiency is improved. The method can be applied to the fieldof nucleic acid in vitro amplification of PCR amplification, loop-mediated isothermal amplification and the like, and has wide application prospects.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for improving the reaction efficiency of nucleic acid in vitro amplification. Background technique [0002] Nucleic acid in vitro amplification is the basis of molecular biology research. With the development of biotechnology, more and more nucleic acid in vitro amplification techniques have emerged, such as polymerase chain reaction (PCR amplification), loop-mediated isothermal amplification Wait. In addition to being used in molecular biology research, nucleic acid in vitro amplification technology is widely used in fields such as precision medicine, food safety testing, and public safety monitoring. Due to the high sensitivity of nucleic acid in vitro amplification technology, if the experimental environment is polluted by aerosols, it is easy to produce false positive results. In addition, since two or more primers are involved in the nucleic acid in vitro ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6848
CPCC12Q1/6848C12Q2527/125
Inventor 李雪玲金彩虹
Owner SHANGHAI NAT ENG RES CENT FORNANOTECH
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