GSM1215608: Anti-CMS_5hmC_Input_Tet1kd; Mus musculus; MeDIP-Seq
Genomic DNA was sheared using Covaris Adaptive Focused Acoustic to the center at 150 bp. DNA fragments were size selected and ligated with methylated adaptors and treated with sodium bisulfite (invitrogen). The extra adapters were removed by AmpurXP (Beckman). The DNA was then denatured for 10 minutes at 95 °C (0.4 M NaOH, 10 mM EDTA), neutralized by the addition of cold 2M ammonium acetate (pH 7.0) for 10 min on ice. Then denatured DNA fragments were incubated with 1 μl anti-CMS antiserum in 1x IP buffer (10 mM sodium phosphate pH 7.0, 140 mM NaCl, 0.05% Triton X-100) for 2 h at 4 °C, and then precipitated with protein G beads. Precipitated DNA was eluted with proteinase K, purified with phenol chloroform, amplified and enriched by 4-6 cycles PCR using Pfu TurboCx hotstart DNA polymerase (Stratagene).  DNA sequencing was carried out using Illumina Genome Analyzer 2 and HiSeq sequencing systems.
GEO
GSM1215608