GSM8181475: CT6A; Mus musculus; OTHER
mice liver tissue suspended 1.0 mL lysis buffer、100 μL protease K（20mg/mL）、100 μL SDS（20%）to 2.0mL centrifuge tube，incubate at 56℃. Transfer 50-100 mg tissue with scalpel blade to the prewarmed centrifuge tube. Incubate at 56℃ for 1-2 h，centrifuge at 12,000×g for 10min in room temperature. Transfer the supernatant toa new 2.0 mL centrifuge tube, add equal volume of Phenol/chloroform/isoamyl alcohol (25:24:1), mix thoroughly by inverting vigorously. Centrifuge at 12,000×g for 10 min in room temperature. Transfer the supernatant to a new 1.5 mL centrifuge tube, taking care not to transfer the intermediate protein layer, add 2/3 of the supernatant volume of isopropanol and 100 μl of sodium acetate, mix well by gently inverting. Transfer to -20°C for 2 hours. Centrifuge at 18213×g for 15 minutes at room temperature and remove the supernatant. Add 1 mL 75% ethanol and wash the precipitation by pipetting. Centrifuge at 18213×g for 3 minutes in room temperature and remove the supernatant. Air dry the precipitation for 3-5 min. Add 20-200 μl TE buffer to resolve. After sample QC, genomic DNA fragment, fragments selection, end Repair, add A and adaptor ligation, PCR, library QC, Circularization, single-stranded circle DNA molecules are replicated via rolling cycle amplification, and a DNA nanoball (DNB) which contain multiple copies of DNA is generated. Sufficient quality DNBs are then loaded into patterned nanoarrays using high-intensity DNA nanochip technique and sequenced through combinatorial Probe-Anchor Synthesis (cPAS). 1. DNA fragmentation 2. Size selection 3. End repair and "A" tailing 4. Adapter ligation 5. Product size selection 6. PCR reaction 7. Library QC Raw data with adapter sequences or low-quality sequences was filtered. We first went through a series of data processing to remove contamination and obtain valid data. This step was completed by SOAPnuke software developed by BGI.