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
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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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