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Quick detection kit for the CYP2C19*2 genetype and detection method thereof

A CYP2C19, 1.CYP2C19 technology, applied in the field of CYP2C19*2 genotype rapid detection kit, can solve the problems of increasing detection operation steps, prolonging detection time, and increasing the risk of disease treatment for patients

Inactive Publication Date: 2015-06-17
重庆京因生物科技有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, when using the reagents currently on the market, the operator needs to prepare the reagents according to the reagent components and instructions in the kit before using them. It takes at least 8 hours for the whole process from blood extraction to DNA test results
Therefore, it is difficult to meet the rapid diagnosis of clinical diseases, which increases the risk of patients' disease treatment

Method used

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  • Quick detection kit for the CYP2C19*2 genetype and detection method thereof
  • Quick detection kit for the CYP2C19*2 genetype and detection method thereof
  • Quick detection kit for the CYP2C19*2 genetype and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0091] 1. Configure the cell lysate:

[0092] Use a 1.5ml centrifuge tube, add 100ulSDS and 18.78ulTriton X-100, then add 881.22ul nuclease-free water to a total volume of 1000ul, make the final concentration of SDS reach 1%, make the final concentration of TritonX-100 reach 2%, That is, 200x cell lysate, then shake and mix well, and store at -4°C.

[0093] The present invention is illustrated by taking the final concentration of Example 2 in the above-mentioned Table 1 and Table 2 as an example, wherein, dNTPs, MgCl 2 , 5x Colorless Reaction Buffer, and DNA polymerase using GoTaq DNA Polymerase were all purchased from Shanghai Promega Biological Products Co., Ltd.

[0094] 2. Prepare 23ul of PCR reaction mixture: the amount of each raw material added is as shown in Table 3:

[0095] table 3

[0096]

[0097]

[0098] Add each raw material according to the amount added in each kit in Table 3 to make a PCR reaction mixture.

[0099] Among them: CYP2C19*2 upstream prim...

Embodiment 1

[0127] The difference from Example 2 lies in the configuration of the cell lysate and the PCR reaction mixture, and the specific operations are as follows:

[0128] 1. Configure the cell lysate:

[0129] Use a 1.5ml centrifuge tube, add 10ul SDS and 1.88ul Triton X-100, then add 988.12ul nuclease-free water to a total volume of 1000ul, make the final concentration of SDS reach 0.1%, and make the final concentration of TritonX-100 reach 2 %, that is, 200x cell lysate, then shake and mix well, and store at -4°C.

[0130] The present invention is illustrated by taking the final concentration of Example 1 in the above Table 1 and Table 2 as an example, wherein, dNTPs, MgCl 2 , 5x Colorless Reaction Buffer, and DNA polymerase using GoTaq DNA Polymerase were all purchased from Shanghai Promega Biological Products Co., Ltd.

[0131] 2. Prepare 23ul of PCR reaction mixture: the amount added is shown in Table 4:

[0132] Table 4

[0133]

[0134]

[0135] Add each raw materia...

Embodiment 4

[0151] The difference from Example 2 lies in the configuration of the cell lysate and the PCR reaction mixture, and the specific operations are as follows:

[0152] 1. Configure the cell lysate:

[0153] Use a 1.5ml centrifuge tube, add 300ul SDS and 56.34ul Triton X-100, then add 643.66ul nuclease-free water to a total volume of 1000ul, make the final concentration of SDS reach 3%, make the final concentration of TritonX-100 reach 6% , that is, 200x cell lysate, then shake and mix well, and store at -4°C.

[0154] The present invention is illustrated by taking the final concentration of Example 4 in the above-mentioned Table 1 and Table 2 as an example, wherein, dNTPs, MgCl 2 , 5x Colorless Reaction Buffer, and DNA polymerase using GoTaq DNA Polymerase were all purchased from Shanghai Promega Biological Products Co., Ltd.

[0155] 2. Prepare 23ul of PCR reaction mixture: the amount added is shown in Table 5:

[0156] table 5

[0157] components Final concentra...

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Abstract

The invention discloses a quick detection kit for the CYP2C19*2 genetype. The quick detection kit comprises a PCR mixed solution, wherein the PCR mixed solution comprises the following raw materials: DNA (deoxyribonucleic acid) polymerases, DNTPs, CYP2C19*2 forward primers, CYP2C19*2 reverse primers, CYP2C19*2 mutant molecular beacons, CYP2C19*2 wild-type molecular beacons, MgC12, 5x Colorless Reaction Buffer and a cell lysis solution. The quick detection kit overcomes the defects that the conventional kit aiming at clinically detecting the CYP2C19*2 genetype and the detection method thereof cannot satisfy quick diagnosis of instructing clinical medication, and the quick detection kit and the detection method thereof provided by the invention can quickly detect the CYP2C19*2 genetype, and reduces patients' medical treatment risk.

Description

Technical field [0001] The present invention is a molecular biology field, which involves a CYP2C19*2 genotype fast detection kit and its detection method. Background technique [0002] Single Nucleotide Polymorphism (SNP) mainly refers to the DNA sequence polymorphism caused by the variation of a single nucleotide at the genome level.In the genomic DNA, any alkali base may mutate, so SNP may be both in the gene sequence or on non -coding sequences other than genes.Overall, there are fewer SNP (CODING SNP, CSNP) in the encoding area, but it is of great significance in the research of hereditary diseases, so the research of CSNP is more concerned. [0003] From the perspective of the influence of creature traits, CSNP can be divided into two types: one is synonymous CSNP, that is, changes in the encoding sequence caused by SNP do not affect the amino acid sequence of protein it translated, mutant alkali and alkali base andThe meaning of the unable mutant base is the same; the othe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q1/6827C12Q2600/156
Inventor 任晓东罗志超
Owner 重庆京因生物科技有限责任公司
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