Application of JAM3 gene to preparation of colorectal cancer diagnosis kit and kit
A diagnostic kit and technology for colorectal cancer, applied in biological testing, biochemical equipment and methods, microbiological measurement/testing, etc., can solve the problem of unclear role of JAM3 in colorectal cancer, and improve the 5-year survival rate , reduce mortality, and have broad application prospects
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Embodiment 1
[0044] Example 1: Detection of differential expression of JAM3 gene in colorectal cancer tissue and corresponding distant normal intestinal tissue by fluorescent quantitative PCR
[0045] 1. Collect 27 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.
[0046] 2. Preparation of RNA samples
[0047] (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 TrizolRNA extraction solution, soak it in 0.1% DEPC water overnight, cut it into pieces with sterilized scissors, and put it into the tissue Grind thoroughly in a homogenizer.
[0048] (2) Extraction: add 500 μL of chlorofo...
Embodiment 2
[0067] Example 2: Detection of differential expression of JAM3 gene in colorectal cancer cell lines and normal intestinal epithelial cell lines by fluorescent quantitative PCR
[0068] 1. Cultivate human colorectal cancer cell lines (SW480, SW620, HCT116 and HT29) and human normal intestinal epithelial cell line NCM460. Mycin 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 was treated with L15 (containing 10% fetal bovine serum), and human intestinal cancer HCT116, SW620 and HT29 cell lines were treated with DMEM (containing 10% fetal bovine serum) at 37°C and 5% CO 2 cultured in an incubator.
[0069] 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 TrizolRNA extraction solution, and gently shake the culture dish left and right to make The ...
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