Human NDRG4/TFPI2 gene methylation detection marker and reagent kit
A detection kit and methylation technology, which are applied in the determination/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problem of low diagnostic accuracy, low specificity and effectiveness, and insufficient early diagnosis of colorectal cancer. Mature, survival time less than 30 months and other issues, to achieve efficient clinical auxiliary diagnosis, high accuracy, increase accuracy and specificity
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[0037] The present invention will be further described in detail below in conjunction with specific embodiments.
[0038] 1. Human NDRG4 / TFPI2 gene methylation quantitative detection system
[0039] Samples: human stool samples, positive quality control products, negative quality control products and standard products (the copy numbers of the plasmid are 100,000, 10,000, 1,000, 100 and 10 respectively);
[0040] Instruments: Lightcycler480, high-speed centrifuge, water bath, PCR instrument, vortex oscillator, ultra-clean workbench;
[0041] The capture primers for ACTB, NDRG4 and TFPI2 genes are as follows:
[0042] Capture probe
Sequence (5’-3’)
ACTB
/ 5AmMC6 / ACCCAGCACACAGTGGCAGACACAGGTTCACAGT
NDRG4
/ 5AmMC6 / TCCCTCGCGCGTGGCTTCCGCCTTCTGCGCGGCTGGGGTGCC
TFPI2
/ 5AmMC6 / CCTGGAGCAGAAAGCCGCGC
[0043] Note: / 5AmMC6 / means probe 5'terminal amino group + carbon 6 molecule
[0044] The methylation primers and probes of ACTB, NDRG4 and TFPI2 genes are as follows:
[0045] Primers, p...
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