Control Mouse Liver Day 152
Whole genome methylation sequencing libraries were prepared using the NEB Enzymatic Methyl-Seq Kit (NEB Cat #E7120L) according to the manufacturers instructions. Briefly, gDNA was acoustically sheared using the PIXUL (Active Motif) to achieve ~300 bp fragments using the suggested protocol: Pulse 50 N/PRF 1 kHz/Burst 20 Hz/Time 36min. 200 ng of sheared gDNA was end repaired and ligated with EM-seq adaptors. Following SPRI bead purification, adaptor-ligated DNA was enzymatically converted. Purified, converted DNA was dual-indexed and amplified by PCR using Q5U polymerase for 4 cycles. Final libraries were size-selected using a 0.85X SPRISelect bead purification, pooled at equimolar ratios, and sequenced on a NextSeq2000 using a 2x150bp strategy.