GSM2127623: HMGB2_ChIPseq_PHE; Rattus norvegicus; ChIP-Seq
NRVMs were fixed (1% formaldehyde, 10 minutes), lysed in lysis buffer (50mM HEPES pH7.5/150mM NaCl/1mM EDTA pH8/1% Triton X-100/0.1% sodium deoxycholate/0.1% SDS, protease inhibitor cocktail tablet [Roche]), sonicated to fragments of 500 bp and diluted in RIPA buffer. DNA-bound protein was immunoprecipitated using anti-HMGB2 (Abcam, ab67282) and precipitated with protein A conjugated magnetic beads (Millipore, LSKMAGA10). Beads were washed (twice in wash buffer: 0.1%SDS/1%Triton X-100/2mM EDTA pH8/150mM NaCl/20mM Tris-HCl pH8; once in 500mM NaCl in wash buffer). DNA was eluted (1%SDS/100mM NaHCO3, 15 minutes, 30°C) and phenol:chloroform purified. Samples were ligated to sequencing adapters with Illumina Paired-End sample prep kit and sequenced on Illumina Genome Analyzer IIx using paired-end sequencing.
GEO Accession
GSM2127623