Rapid PCR amplification kit as well as application method thereof

An application method and kit technology, applied in the field of genetic engineering, can solve the problems of STR typing failure, low DNA detection rate, low reaction sensitivity, etc., and achieve the effects of rapid amplification, saving money, and low error rate

Inactive Publication Date: 2019-12-03
CYTTEL BIOSCI BEIJING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the little DNA residue at the crime scene and the traces of forensic evidence DNA collected are extremely small, the reaction sensitivity of conventional PCR amplification methods is low, it is difficult to capture effective DNA information, and the amplification effect is poor, which directly leads to subsequent DNA testing. The detection rate is low, STR typing fails, and criminal individuals cannot be identified

Method used

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  • Rapid PCR amplification kit as well as application method thereof
  • Rapid PCR amplification kit as well as application method thereof
  • Rapid PCR amplification kit as well as application method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Using the rapid PCR amplification kit, reaction system and reaction conditions of the present invention to amplify the internal reference gene A segment in human genomic DNA, and take appropriate amount of mouse heart, liver, spleen, lung, kidney, brain, muscle, toe, mouse Nucleic acid DNA PCR amplification experiment for tail, bone, hair tissue, see figure 1 The amplified agarose gel electrophoresis test results shown, in which lane 1 is the DNA marker, and lanes 2 to 12 are the mouse heart, liver, spleen, lung, kidney, brain, muscle, toe, mouse tail, and bone respectively 1. Nucleic acid DNA amplification result of hair tissue, 13 swimming lanes are human genome DNA amplification results, 14 swimming lanes are negative control (i.e. ddH 2 O is the template).

Embodiment 2

[0042] Using the rapid PCR amplification kit, reaction system and reaction conditions of the present invention to amplify the internal reference gene B segment in human genomic DNA, and take an appropriate amount of mouse heart, liver, spleen, lung, kidney, brain, muscle, toe, mouse Nucleic acid DNA extraction and PCR amplification experiments for tail, bone, hair tissue, see figure 2The amplified agarose gel electrophoresis test results shown, in which lane 1 is the DNA marker, and lanes 2 to 12 are the mouse heart, liver, spleen, lung, kidney, brain, muscle, toe, mouse tail, and bone respectively 1. Nucleic acid DNA amplification result of hair tissue, 13 swimming lanes are human genome DNA amplification results, 14 swimming lanes are negative control (i.e. ddH 2 O is the template).

[0043] The rapid PCR amplification kit of the present invention and its application method, compared with the prior art PCR amplification kit, uses Phire hot start II DNA polymerase in combin...

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PUM

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Abstract

The invention provides a rapid PCR amplification kit as well as an application method thereof. The kit comprises DNA polymerase, dNTPs, internal reference gene primers and buffer fluid lines, whereinthe internal reference gene primers comprise an internal parameter A fragment and an internal parameter B fragment, the internal parameter A fragment comprises an upstream primer F and a downstream primer R, and the internal parameter B fragment comprises an upstream primer F and a downstream primer R. According to the rapid PCR amplification kit as well as the application method thereof, combinedwith Phire heat to start IIDNA polymerase, amplification of longer fragment targets can be realized, mistake rate is lower, a PCR enhancer is added into the kit, a high-level structure of a nucleotide can be opened, the kit has the characteristics by being convenient, saving money and improving efficiency, and rapid amplification of trace DNA samples is realized through one-step PCR detection.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a rapid PCR amplification kit and an application method thereof. Background technique [0002] PCR is the abbreviation of polymerase chain reaction. It is a molecular biology technique used to amplify specific DNA fragments. Its research has a profound impact on various fields of biology, and its relationship with medicine, agriculture, bioengineering, etc. Very closely, for example, PCR technology involves new ways of breeding in agriculture, medical detection and treatment of genetic diseases, emerging industries based on genetic engineering, forensic evidence identification, paternity testing, etc. Rapid PCR includes two parts: rapid nucleic acid extraction and PCR amplification. The classic PCR amplification kit includes PCR buffer system, dNTPs, DNA polymerase, etc. In order to prepare the PCR reaction solution in the existing kits, different reagents are repeat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/686
CPCC12Q1/686C12Q2521/101C12Q2527/125C12Q2545/101
Inventor 孙晓娇张文平
Owner CYTTEL BIOSCI BEIJING
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