Polymerase chain reaction enhancing method

A chain reaction and polymerase technology, applied in the biological field, can solve problems such as incomplete PCR amplification, and achieve the effect of improving sensitivity and efficiency

Inactive Publication Date: 2017-04-26
SANSURE BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the amplification of some specific nucleic acid fragment regions, the design position of primers is limited; or in multiple PCR reactions, the optimization of PCR buffer solution composition, annealing temperature and other conditions is limited, etc., which will lead to poor PCR amplification. perfect

Method used

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  • Polymerase chain reaction enhancing method
  • Polymerase chain reaction enhancing method
  • Polymerase chain reaction enhancing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] The DNA extracted from the oropharyngeal swab samples of 8 respiratory adenovirus (ADV) infected patients was used as the sample to be tested, one was tested with a PCR reaction system without an enhancer as a control, and the other was tested with sorbitol and The PCR reaction system detection of trehalose was used as the experimental group, figure 2 and image 3 Shown:

[0078] Ct value statistics of control group and experimental group:

[0079]

[0080] Visible, the present invention provides the method for polymerase chain reaction enhancement, adopt the mixture that adds 0.2M sorbitol and 0.25M trehalose as PCR enhancer and can significantly improve the efficiency of 8 respiratory tract adenovirus (ADV) DNA nucleic acid amplification reactions and and / or yield, thereby improving the sensitivity of nucleic acid detection using nucleic acid amplification reactions.

Embodiment 2

[0082] Use the RNA extracted from the plasma samples of 7 human immunodeficiency virus (HIV) infected patients as the test sample, one is tested with the PCR reaction system without adding enhancer as the control, and the other is added with sorbitol and trehalose The detection of the PCR reaction system is used as the experimental group, and the result Figure 4 and Figure 5 Shown: Ct value statistics of control group and experimental group:

[0083]

[0084] Visible, the present invention provides the method for polymerase chain reaction enhancement, adopt the mixture of 0.1M sorbitol and 0.25M trehalose as PCR enhancer and can significantly improve the efficiency of 7 human immunodeficiency virus (HIV) RNA nucleic acid amplification reactions and and / or yield, thereby increasing the sensitivity of nucleic acid detection using nucleic acid amplification reactions

Embodiment 3

[0086] Detect the α-globin gene of human peripheral blood sample DNA, and analyze the results by agarose gel electrophoresis as follows: Figure 6 Shown:

[0087] Among them, α-thalassemia αα / αα wild-type amplification results (1.7kb) 3: DNA Markers; 1: Experimental group (addition of 0.2M sorbitol and 0.25M trehalose);

[0088] 2 control group (not added);

[0089] It can be seen that the present invention provides a method for enhancing the polymerase chain reaction, using a mixture of 0.2M sorbitol and 0.25M trehalose as a PCR enhancer can significantly improve the efficiency of the amplification reaction of α-globin nucleic acid in human peripheral blood sample DNA and / or yield, thereby improving the sensitivity of nucleic acid detection using nucleic acid amplification reactions.

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Abstract

The invention discloses a polymerase chain reaction enhancing method. The polymerase chain reaction enhancing method includes the steps of adding a PCR (polymerase chain reaction) enhancer into a PCR amplification system so as to obtain an enhanced PCR amplification system, wherein the PCR enhancer refers to a mixture of hexanehexol and trehalose; setting program parameters of the enhanced PCR amplification system, and conducting amplification test so as to obtain an amplified product; testing the amplified product on a fluorescence quantitative PCR amplification instrument. The polymerase chain reaction enhancing method has the advantages that since the mixture of the hexanehexol and the trehalose is used as the PCR enhancer, nucleic acid amplification reaction efficiency and/or yield can be remarkably improved and/or increased, and the nucleic acid sensitivity detected by nucleic acid amplification is improved.

Description

technical field [0001] The invention relates to the technical field of biotechnology, in particular to a method for enhancing polymerase chain reaction. Background technique [0002] Polymerase chain reaction (polymerase chain reaction; PCR) is a molecular biology technique used to amplify specific nucleic acid fragments (DNA / RNA). The characteristic is that it can greatly increase the trace amount of nucleic acid. General PCR mainly adopts at least two kinds of oligonucleotide primers, upstream and downstream, combined with the selected nucleic acid template (such as DNA or RNA), and under the action of DNA polymerase (such as Taq DNA polymerase, Tth DNA polymerase), target Nucleic acid template fragments are amplified. The above two oligonucleotide primers can be purified from restriction digests by conventional methods, or the above primers can be produced synthetically. Primers are preferably single-stranded for the most efficient amplification, but primers may also b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12Q1/68
CPCC12Q1/6806
Inventor 陈晓亮周雷刘佳邓中平戴立忠
Owner SANSURE BIOTECH INC
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