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Lysate and kit for carrying out PCR on blood and oral cavity swab

A lysing solution and kit technology, applied in the field of lysing solution and kits, can solve the problems of low extraction efficiency, high extraction cost and shortage, and achieve the effects of saving time and cost, strong anti-interference ability and low cost.

Inactive Publication Date: 2019-03-01
广州海思医疗科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are some shortcomings in these methods: 1) the extraction cost of the first two is high, and the extraction efficiency is low; 2) the traditional phenol-chloroform extraction method has the problems of cumbersome extraction steps and insufficient purity
In addition, during the nucleic acid extraction process, due to the influence of the operator's proficiency and experimental conditions, DNA contamination or insufficient extraction often occurs, which will lead to the failure of subsequent research or detection.

Method used

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  • Lysate and kit for carrying out PCR on blood and oral cavity swab
  • Lysate and kit for carrying out PCR on blood and oral cavity swab
  • Lysate and kit for carrying out PCR on blood and oral cavity swab

Examples

Experimental program
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Effect test

Embodiment 1

[0033] This embodiment provides a lysate, which consists of the following components: Tris-Hcl 67mM, ammonium sulfate 16.6mM, Mgcl 2 ·6H 2 O 6.7mM, EDTA 6.7μM, SDS 1.7μM and proteinase K 50μg / ml, the pH value of Tris-Hcl was 8.8.

Embodiment 2

[0035] This embodiment provides a lysate, which consists of the following components: Tris-Hcl 70mM, ammonium sulfate 17mM, Mgcl 2 ·6H 2 O 7mM, EDTA 7μM, SDS 2.5μM and proteinase K 55μg / ml, the pH value of Tris-HCl was 9.

Embodiment 3

[0037] This embodiment provides a kind of lysate, and it is made up of the component of following concentration: Tris-Hcl 60mM, ammonium sulfate 16mM, Mgcl 2 ·6H 2 O 6mM, EDTA 6μM, SDS1.5μM and proteinase K 30μg / ml, the pH value of Tris-Hcl was 8.2.

[0038] The lysate of Example 1 was used for the detection of the following samples, and the PCR amplification instrument used in the detection was Tianlong TL988.

[0039] Take whole blood samples from normal people, and perform real-time fluorescent quantitative PCR detection on the SNP sites

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Abstract

The invention discloses lysate and kit for carrying out a PCR on a blood and oral cavity swab, and belongs to the technical field of molecular diagnosis. The kit comprises the lysate. The lysate is prepared from 60-70 mM of Tris-Hcl, 16-17 mM of ammonium sulfate, 6-7 mM of Mgcl2 6H2O, 6-7 micron M of EDTA, 1.5-2.5 micron M of SDS and 30-55 micrograms / ml of protease K, wherein the pH value of Tris-Hcl is 8.2-9. The blood and oral cavity swab treated with the lysate can be directly subjected to the PCR, and an existing detection technology is greatly optimized.

Description

technical field [0001] The invention belongs to the technical field of molecular diagnosis, and in particular relates to a lysate and a kit for performing PCR on blood and buccal swabs. Background technique [0002] PCR, or Polymerase Chain Reaction (Polymerase Chain Reaction), is an enzymatic synthesis reaction technology that relies on DNA polymerase in the presence of template DNA, primers and four deoxyribonucleotides. This technology has been widely used in the fields of microbiology, genetics and forensics since it was invented in 1958 by Kary Mullis and colleagues in the human genetics laboratory of Cetus Corporation in the United States. [0003] Real-time fluorescent quantitative PCR technology (qPCR) was introduced by Applied Biosystems in 1996. This technology refers to adding fluorescent groups or fluorescent dyes to the PCR reaction system, using the accumulation of fluorescent signals to monitor the entire PCR process in real time, and finally drawing a standar...

Claims

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Application Information

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IPC IPC(8): C12Q1/686
CPCC12Q1/686
Inventor 夏昊强姚啟聪江伟钊王芳周煌凯刘倩关丽雅邓美英程祖福
Owner 广州海思医疗科技有限公司
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