Use of the product for detecting the methylation site of the ndrg4 gene in the preparation of a product for the early detection of colorectal cancer
A technology for early detection of colorectal cancer, applied in the field of biotechnology and disease detection, can solve problems such as inactivation of tumor suppressor genes
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Embodiment 1
[0389] Example 1. Using TaKaRa MiniBEST FFPE DNA Extraction Kit to Extract Genomic DNA from FFPE Samples
[0390] Specific steps are as follows:
[0391] Use a sterilized scalpel to scrape 30 mg of paraffin section tissue to remove excess paraffin;
[0392] Put the paraffin section tissue into a 1.5mL centrifuge tube, add 500μL Buffer DP, mix well, place in a water bath at 80°C for 1 minute, vortex for 10 seconds while hot, add 180μL Buffer GL, and vortex;
[0393] Then centrifuge at 12000rpm at room temperature for 1 minute, the solution forms two layers, the upper oil phase, the lower water phase, add 20 μL Proteinase K, 20 mg / mL and 10 μL RNase, 10 mg / mL to the lower water phase, pipette to mix, and then 56 ° C water bath After 1 hour, place the treated sample in a 90°C water bath for 30 minutes, and cool to room temperature;
[0394] Add 200 μL Buffer GB and 200 μL 100% ethanol to the processed sample, and vortex for 10 seconds.
[0395] Then centrifuge at room temperat...
Embodiment 2
[0405] Example 2. Genomic DNA Extraction from Feces Samples
[0406] Take a stool sample (4-6g) and add 40mL of lysate, vortex, mix thoroughly and incubate at 50°C for 16 hours;
[0407] After incubation, centrifuge at 5000rpm for 10min. Pay attention to weighing and balance before centrifugation. After centrifugation, carefully take out the centrifuge tube and do not shake violently;
[0408] Pipette 9 mL of supernatant into a new 50 mL centrifuge tube, then add 1 mL of extraction auxiliary solution, 60 μL of magnetic bead solution, and 10 mL of isopropanol, vortex for 10 seconds, incubate at 65°C for 20 minutes, and mix up and down every 5 minutes during the incubation period. Even once;
[0409] After the incubation, place the 50mL centrifuge tube on the magnetic stand and let it stand for 3 minutes. After the magnetic beads are fully adsorbed on the tube wall, discard the waste liquid;
[0410] Take the centrifuge tube out of the magnetic stand, add 12mL washing solution...
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