Cycli-dependent kinase 6 (CDK6) gene exon genetic groups used for cancer diagnosis and diagnosis kit
A technology of cancer diagnosis and kit, which is applied in the direction of recombinant DNA technology, microbial measurement/testing, DNA/RNA fragments, etc., and can solve problems such as differences in overexpression levels
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Embodiment 1
[0095] Example 1: Fluorescent quantitative PCR method to detect the amplification of CDK6 gene Exon 1 in different cancer tissues (intestinal cancer and breast cancer) and corresponding normal tissues adjacent to cancer (intestine and breast)
[0096] 1. Collect 1 pair of colon cancer and breast cancer tissues and corresponding paracancerous bowel and breast normal tissues. All tissue samples are provided by the Specimen Bank of the First Affiliated Hospital of Xiamen University. Each patient specimen has a clear diagnosis record , and with the consent of the ethics committee, the specimens were collected and frozen in liquid nitrogen.
[0097] 2. DNA sample preparation
[0098] (1) Sample pretreatment: Take out the sample from the liquid nitrogen tank, put 30mg of the sample into a 2ml sterile EP tube, add 600μL of DNA extraction solution and 20mg / ml proteinase K, fully cut it into pieces with sterilized scissors, and then 65 Incubate for 1 hour at °C.
[0099] (2) Centrifu...
Embodiment 2
[0110] Example 2: Fluorescent quantitative PCR method to detect the amplification of CDK6 gene Exon 1-8 in 5 pairs of breast cancer tissues and corresponding adjacent normal tissues
[0111] 1. Collect 5 pairs of breast cancer tissue and corresponding normal adjacent tissue samples. All 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 has been approved by the ethics committee , collected specimens and frozen in liquid nitrogen.
[0112] 2. DNA extraction and fluorescent quantitative PCR methods are the same as in Example 1.
[0113] 3. Data analysis: the experiments were repeated 3 times, and relative quantification 2 -△△Ct (△△Ct=(cancer tissue CDK6Exon1-8Ct value-cancer tissue Exon1-8GAPDH Ct value)-(normal tissue CDK6Exon1-8Ct value-normal tissue Exon1-8GAPDH Ct value)) method for statistical analysis, GAPDH as an internal reference gene, The data were analyzed using...
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