Application of pcdh18 gene in preparation of colorectal cancer diagnostic kit and kit
1. The technology of PCDH18 and diagnostic kits is applied in the direction of biological testing, biochemical equipment and methods, and the measurement/testing of microorganisms, which can solve the problems that no research reveals the relationship between the development of colorectal cancer, etc., and improve the 5-year survival rate, mortality reduction, and timely diagnosis
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Embodiment 1
[0052] Example 1: Fluorescent quantitative PCR method to detect the differential expression of PCDH18 gene in colorectal cancer tissue and corresponding distant normal intestinal tissue
[0053] 1. Collect 25 pairs of colorectal cancer tissue and corresponding distant normal intestinal tissue samples. All colorectal cancer tissue samples are provided by the Specimen Bank of the First Affiliated Hospital of Xiamen University. Each patient sample has a clear diagnosis record, and passed With the approval of the ethics committee, the specimens were collected and frozen in liquid nitrogen.
[0054] 2. Preparation of RNA samples
[0055] (1) Pretreatment of the sample: Take the sample out of the liquid nitrogen tank, put the sample into a 2ml sterile EP tube, add 1ml Trizol RNA extraction solution, soak it in 0.1% DEPC water overnight and cut it into pieces with sterilized scissors. Grind thoroughly in a tissue homogenizer.
[0056] (2) Extraction: add 500 μL of chloroform, vigor...
Embodiment 2
[0075] Example 2: Fluorescent quantitative PCR method to detect the differential expression of PCDH18 gene in colorectal cancer cell lines and normal intestinal epithelial cell lines
[0076] 1. Cultivate human colorectal cancer cell lines (SW480, SW620, HCT116 and HT29) and human normal intestinal epithelial cell line NCM460. Mycin each 100U / mL in DMEM medium, at 37°C and 5% CO 2 Cultured in an incubator, the culture medium was changed every 2-3 days and subcultured. Human intestinal cancer SW480 uses L15 medium (containing 10% fetal bovine serum), and human intestinal cancer HCT116, SW620 and HT29 cell lines adopts DMEM medium (containing 10% fetal bovine serum). 2 cultured in an incubator.
[0077] 2. When the confluence of the cells in the 60mm culture dish reaches 80%, discard the culture medium, add 1mL sterile PBS to wash, discard the PBS and repeat once, add 1mL Trizol RNA extraction solution, and gently shake the culture dish left and right , so that all the Trizol...
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