Fluorescent quantitative PCR diagnostic kit for rapidly detecting HER-2 mRNA
A HER-2, fluorescence quantitative technology, applied in fluorescence/phosphorescence, material excitation analysis, etc., to achieve simple and safe operation, low false positives, and good specificity
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[0017] The present invention will be further described below in conjunction with a specific embodiment. This embodiment is only used to illustrate the present invention and is not intended to limit the scope of protection claimed by the present invention.
[0018] (1) The kit of the present invention consists of the following:
[0019] ①Trizol
[0020] ② First strand cDNA synthesis kit (RT-PCR)
[0021] ③ Primers: including the target gene HER-2 and the internal reference gene β-actin, as follows:
[0022] ④ Upstream: 5′-GACCGGAGGCTGACCAGT-3′;
[0023] ⑤Downstream: 5′-GGCAAGGCTGGCATGCGG-3′;
[0024] ⑥Beta-actin gene primer sequence:
[0025] ⑦Upstream: 5'-GAAGATGGTGATGGGATTTC-3';
[0026] Downstream: 5'-GAAGGTGAAGGTCGGAGTC-3';
[0027] ⑧Fluorescent probe: FAM 5'-GGCCCGCTGCCCCAGCGGTG-3'TAMRA;
[0028] FAM5'-ACCACCACGGCCGAGCGG-3'TAMRA
[0029] ⑨PCR reaction solution: 1×PCR buffer, 0.3-0.5pmol / μL primer, 0.1-0.3pmol / μL probe, 2.5-4.0mM Mg2+, 1-2U Taq enzyme, 0.2-0.4mM dN...
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