Unicell simplified representative bisulfite sequencing method and kit
A bisulfite and single-cell technology, applied in biochemical equipment and methods, chemical libraries, combinatorial chemistry, etc., can solve the problem of low coverage of CpG islands
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[0071] Example 1 Using mouse oocytes as materials to construct scRRBS (single-cell simplified representative bisulfite) sequencing library
[0072] (1) lysed cells
[0073] Isolate a single mouse oocyte and place it in a 1.5mL sampling tube with a volume of about 1μL; add 1μL of ddH to the negative control tube 2 O. Without transferring the tube, directly add 5 μL LysisBuffer, 1 μL Protease, and 1 μL λDNA to the cells in a total of 7 μL, the total volume is 8 μL, and wrap the parafilm to seal. Submerge the entire small tube in a water bath at 50°C for 3 hours, and then inactivate Protease at 70°C for 15 minutes. A mixed solution containing mouse oocyte genomic DNA was obtained.
[0074] (2) MspI digestion
[0075] Digest mouse oocyte genomic DNA according to the following system, wherein the volume of the mixture (mouse oocyte gDNA) obtained from step (1) pretreatment is 8 μL (already containing λDNA),
[0076]
[0077] Submerge the entire vial in a water bath at 37°C ...
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