Detection reagent for PIEZO2 gene in human gastrointestinal tumor
A detection reagent and gastrointestinal technology, applied in the field of PIEZO2 gene detection reagents in human gastrointestinal tumors, can solve the problems of complex detection process, delay in diagnosis and treatment of patients, long detection time, etc., to achieve flexible and fast detection, targeted Strong and efficient effect
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Embodiment 1
[0010] human gastrointestinal tumors PIEZO2 Gene detection reagent, the detection reagent is a pair of specific primers, the nucleotide sequence of the upstream primer is: 5'- GGAGTTAGGC GGGAGTATAG TAC -3', the nucleotide sequence of the downstream primer is: 5'- TTCTTCAAAA ATAACTAATA CCGAA -3'. is directed at PIEZO2 The fluorescent quantitative methylation-specific amplification upstream primers and fluorescent quantitative methylation-specific amplification downstream primers designed for the promoter region of the gene were confirmed by our extensive screening tests. The primers were provided by Shanghai Shengong Bioengineering Co., Ltd. synthesis.
Embodiment 2
[0012] Utilize a pair of primers of embodiment 1 to carry out case detection, specific detection process is as follows:
[0013] 1. DNA extraction: We collected 5 cases of gastric cancer paraffin samples and corresponding paracancerous paraffin samples from Ningbo Third People's Hospital, and extracted DNA samples with QIAamp DNA FFPE Tissue Kit (purchased from Qiagen, Hilden, Germany). We collected 5 cases of colorectal cancer paraffin samples and corresponding paracancerous tissue paraffin samples from Ningbo Third People's Hospital, and extracted them in the same way to obtain DNA samples. DNA samples require that the ratio of DNA absorbance (A) (A260 / A280) is about 1.80 .
[0014] 2. Methylation of DNA samples: EZ DNA methylation kit (purchased from Zymo, USA) was used to convert the sample DNA to bisulfite, and after the DNA sample was treated with bisulfite, methylation could occur. The cytosine (C) base remains unchanged, and the unmethylated C is converted into uracil...
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