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Molecular index, kit and method for sample quality control before lung cancer EGFR gene Sanger sequencing

A kit and genetic technology, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of insufficient progress and achieve the effect of reducing time, improving success rate and detection efficiency

Inactive Publication Date: 2016-08-17
广州迈景基因医学科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Common CT scan screening is effective in reducing lung cancer mortality by about 20%, but its progress is far less than that of other common malignancies such as breast, prostate and colon cancer

Method used

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  • Molecular index, kit and method for sample quality control before lung cancer EGFR gene Sanger sequencing
  • Molecular index, kit and method for sample quality control before lung cancer EGFR gene Sanger sequencing
  • Molecular index, kit and method for sample quality control before lung cancer EGFR gene Sanger sequencing

Examples

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Embodiment 1

[0030] Embodiment 1: molecular index

[0031] A molecular index used for quality control of samples of lung cancer EGFR gene before Sanger sequencing according to an embodiment of the present invention includes the following genes: β-actin, PRPH, GABARAPL2, ACTG1, NDUFA7, UQCRC1, MYC and MIF.

Embodiment 2

[0032] Embodiment 2: Kit

[0033] A kit for quality control of samples before Sanger sequencing of the lung cancer EGFR gene according to an embodiment of the present invention, the kit contains primer pairs for the following genes: β-actin, PRPH, GABARAPL2, ACTG1, NDUFA7, UQCRC1, MYC and MIF; the kit also contains 10×PCR buffer solution, 10mmol / L dNTP, more than 50ng cDNA, 0.8μg / μL BSA, rTaq enzyme and ultrapure water; PCR buffer solution, dNTP, gene primer pair, cDNA, BSA, The volume ratio of rTaq enzyme and ultrapure water is: 3:0.8:3:1:2.5:0.4:19.3.

[0034] Wherein, the sense primer of the β-actin is shown in SEQ ID NO: 1, the antisense primer of β-actin is shown in SEQ ID NO: 2; the sense primer of the PRPH is shown in SEQ ID NO: 3, PRPH The antisense primer of described GABARAPL2 is shown in SEQ ID NO:4; The sense primer of described GABARAPL2 is shown in SEQ ID NO:5, and the antisense primer of GABARAPL2 is shown in SEQ ID NO:6; The sense primer of described ACTG1 is ...

Embodiment 3

[0040] Example 3: Method for sample quality control before Sanger sequencing of lung cancer EGFR gene

[0041] A method for quality control of samples before Sanger sequencing of lung cancer EGFR gene according to an embodiment of the present invention, comprising the following steps:

[0042] (1) Fix the lung cancer tissue samples with 10% neutral formalin, identify the tumor area of ​​the lung cancer tissue samples by HE staining, and extract the DNA of the lung cancer tissue samples with QIAgen_FFPE_DNA_kit;

[0043] (2) Select a lung cancer tissue sample with a fixed time of 12-24 hours, a tumor area with more than 100 tumor cells, and an extracted DNA concentration of 1.5-2.0ng / μL; use the DNA of the selected sample as a template, and use β-actin The primer pairs of , PRPH, GABARAPL2, ACTG1, NDUFA7, UQCRC1, MYC and MIF are primers for PCR amplification, the PCR reaction system is as the kit described in Example 2, see Table 2 for details; the PCR reaction conditions are s...

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Abstract

The invention relates to a molecular index, kit and method for sample quality control before lung cancer EGFR gene Sanger sequencing. The molecular index for the sample quality control before Sanger sequencing comprises the following genes of beta-actin, PRPH, GABARAPL2, ACTG1, NDUFA7, UQCRC1, MYC and MIF; the kit comprises primer pairs containing the above-mentioned genes. The eight genes of beta-actin, PRPH, GABARAPL2, ACTG1, NDUFA7, UQCRC1, MYC and MIF serve as the molecular index for the sample quality control before lung cancer tissue Sanger sequencing, the EGFR gene Sanger sequencing success rate can be increased, the detection efficiency of the EGFR gene can be improved, and unnecessary time, manpower and funds can be reduced.

Description

technical field [0001] The invention relates to a molecular index, a kit and a method for sample quality control, in particular to a molecular index, a kit and a method for sample quality control before EGFR gene Sanger sequencing of lung cancer. Background technique [0002] Lung cancer has become a common malignant tumor in all countries in the world. The "two firsts" of lung cancer are recognized at home and abroad, that is, "the first in incidence rate" and "the first in mortality". In the past 50 years, many countries have reported that the incidence and mortality of lung cancer have increased significantly. The incidence and mortality of lung cancer in men account for the first of all malignant tumors, and the incidence and mortality of lung cancer occupy the second place in women. It has become the malignant tumor that threatens human health and life the most. Common CT scan screening is effective in reducing lung cancer mortality by about 20%, but its progress is fa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/156
Inventor 柯尊富王连唐杨冬成
Owner 广州迈景基因医学科技有限公司