GSM4257387: Mouse_liver_KAS-Input.rep1; Mus musculus; OTHER
N3-kethoxal labeling genomic DNA(gDNA) in mouse tissue was isolated. 1 µg genomic DNA was then suspended in 95 µL DNA elution buffer supplemented with 5 µL 20 mM DBCO-PEG4-biotin (DMSO solution, Sigma, 760749), and incubated at 37 ˚C for 1.5 h with gentle shake. 5 µL RNase A (Thermo, 12091039) was added into the reaction mixture followed by an incubation at 37 ˚C for 5 min. Biotinylated gDNA was then recovered by DNA Clean & Concentrator-5 kit (Zymo). gDNA was suspended into 100 µL water and subjected to sonication by using Bioruptor Pico at 30s-on/30s-off setting for 30 cycles. 5% of the fragmented DNA was saved as input, and the rest 95% was used to enrich biotin-tagged DNA by incubating with 10 µL pre-washed Dynabeads MyOne Streptavidin C1 (Thermo, 65001) at room temperature for 15 min. The beads were washed and DNA was eluted by heating the beads in 15 µL H2O at 95 ˚C for 10 min. Eluted DNA and its corresponding input were used for library construction by using Accel-NGS Methyl-seq DNA library kit (Swift, 30024).
GEO
GSM4257387