GSM8620906: aceA; Mus musculus; OTHER
DNA extraction was performed following the protocol of Wizard Genomic DNA Purification Kit (Promega). 1. Add 600 μl of Nuclei Lysis Solution, and pipet until no visible cell clumps remain. 2. Add 3 μl of RNase Solution to the nuclear lysate and mix the sample. Incubate the mixture for 30 minutes at 37 °C. 3. Add 200 μl of Protein Precipitation Solution and vortex at high speed for 20 seconds. Chill sample on ice for 5 minutes. 4. Centrifuge for 10 minutes. Carefully remove the supernatant and transfer it to a clean 1.5 ml tube. 5. Add 600 μl isopropanol and gently mix the solution until the white thread-like strands of DNA form a visible mass. 6. Centrifuge for 15 minutes and carefully decant the supernatant. 7. Add 600 μl of 70% ethanol, and gently invert the tube several times to wash the DNA. Centrifuge for 5 minutes. 8. Carefully decant the ethanol and air-dry for 10 minutes. 9. Add 100 μl of DNA Rehydration Solution and incubate at 37 °C for 30 minutes. Library was constructed following the protocol of EpiArt DNA Methylation Library Kit for Illumina V3 (Vazyme). 1. Denature the input DNA into single-stranded DNA at 95 °C for 2 minutes and chill sample on ice for 2 minutes. 2. Add the 3' adapter to the 3' end of the single-stranded DNA at 37 °C for 15 minutes. 3. Extend single-stranded DNA to double-stranded DNA by extension primers. 4. Purify the reaction products using 1.2 × VAHTS DNA Clean Beads. 5. Add the 5' adapter to the 5' end of the double-stranded DNA at 25 °C for 15 minutes. 6. Purify the reaction products using 1 × VAHTS DNA Clean Beads. 7. Amplify DNA library by index primers for 10 cycles. 8. Purify the reaction products using 0.85 × VAHTS DNA Clean Beads. 9. Determine the library concentration by Qubit.