GSM8381337: WT2; Mus musculus; ChIP-Seq
muscle from mice was collected and instantly frozen in liquid nitrogen for conservation and transportation, and anti-H3K9me3 antibody (#13969, Cell Signaling) was used to map its binding sites in the 384 genome. DNA was purified, genomic fragments with adapters at both ends were amplified by PCR, sequenced and mapped to the genome for further analysis. To amplify libraries, 21 µL DNA was mixed with 2 µL of a universal i5 and a uniquely barcoded i7 primer33, using a different barcode for each sample. A volume of 25 µL NEBNext HiFi 2× PCR Master mix was added and mixed. The sample was placed in aThermocycler with aheated lid using the following cycling conditions: 72 °C for 5 min (gap filling); 98 °C for 30 s; 14 cycles of 98 °C for 10 s and 63 °C for 30 s; final extension at 72 °C for 1 min and hold at 8 °C. Post-PCR clean-up was performed by adding 1.1× volume of Ampure XP beads (Beckman Counter), and libraries were incubated with beads for 15 min at RT, washed twice gently in 80% ethanol, and eluted in 30 µL 10 mM Tris pH 8.0.