GSM4879519: GR_2D_no training_Corticosterone; Rattus norvegicus; ChIP-Seq
The snap-frozen, disected hippocampi were fixated with 1% formaldehyde and homogenized in Jiang buffer using a glass douncer (Kimble-Chase). Chromatin of 4 hippocampi of 2 rats was pooled and suspended in NP-50 containing lysis buffer for fragmentation by sonication. Input aliquots were taken and chromatin samples were split in two for paired GR and pCREB ChIP, using anti-GR antibody H-300 (sc-8992X, Santra Cruz) and anti-phospho-CREB Ser133 antibody (17-10131, Millipore), respectively. Transcription factor bound DNA was collected, decrosslinked and purified by phenolization. Adapters (Agilent) were ligated and samples were subjected to 15 rounds of PCR for DNA library preparation (KAPA Biosystems). Single-end sequencing was performed on a Illumina HiSeq 2500 at High Output. Two runs were performed to obtain sufficient data, first 51bp and secondly 65bp sequencing.