Ribonucleotide reductase subunit M1 gene expression fluorescence quantitative PCR detection kit
A detection kit and fluorescence quantitative technology, applied in the field of molecular biology, can solve the problems of gemcitabine drug resistance, drug inactivity, increased expression, etc., to achieve improved detection sensitivity and specificity, good specificity, and low false positives Effect
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Embodiment 1
[0042] Example 1: The RRM1 gene expression fluorescent quantitative PCR kit of the present invention detects two genotypes of A / C and A / A at the -37 site of the RRM1 gene in tissues of patients with non-small cell lung cancer.
[0043] One probe and one pair of common primers can be designed to specifically detect the A / C and A / A genes at the -37 site.
[0044] The forward primer sequence is:
[0045] SEQ ID NO: 1 (5'-AAGAG CAGCG TGCCA GAGAT-3');
[0046] The reverse primer sequence is:
[0047] SEQ ID NO:2 (5'-ACACA TCAAA GACCA GTCCT GATTA G-3');
[0048] The fluorescent probe sequence is:
[0049] SEQ ID NO: 3 (FAM-TTGC TCTTT GGATT CCGGA TCTCT TCA-BHQ1). The 5' end of the fluorescent probe is labeled with FAM (fluorescence reporter group), and the 3' end is labeled with BHQ1 (fluorescent quencher group).
[0050] Then optimize the reaction system for FQ-PCR detection: the reaction system is 15 μl, forward and reverse primers 0.15 μl (20 μΜ), each probe 0.2 μl (20 μΜ), t...
Embodiment 2
[0053] Example 2: The RRM1 gene expression fluorescent quantitative PCR kit of the present invention detects two genotypes of C / T and C / A at the -524 site of the RRM1 gene in tissues of patients with non-small cell lung cancer.
[0054] One probe and one pair of common primers can be designed to specifically detect the -524 site to detect C / T and C / A genes.
[0055] The forward primer sequence is:
[0056] SEQ ID NO: 1 (5'-AAGAG CAGCG TGCCA GAGAT-3');
[0057] The reverse primer sequence is:
[0058] SEQ ID NO:2 (5'-ACACA TCAAA GACCA GTCCT GATTA G-3');
[0059] The fluorescent probe sequence is:
[0060] SEQ ID NO: 3 (FAM-TTGC TCTTT GGATT CCGGA TCTCT TCA-BHQ1). The 5' end of the fluorescent probe is labeled with FAM (fluorescence reporter group), and the 3' end is labeled with BHQ1 (fluorescent quencher group).
[0061] Then optimize the reaction system for FQ-PCR detection: the reaction system is 15 μl, forward and reverse primers 0.15 μl (20 μΜ), each probe 0.2 μl (20 μ...
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