title: GSM1697674: n5-iPSC.input.ChIP-Seq; Homo sapiens; ChIP-Seq
library_construction_protocol: For RNA-Seq, total RNA was isolated from naïve iPSC lines and primed PSC lines using Trizol reagent (Invitrogen) according to the manufacturer’s instructions. The RNA sequencing libraries were constructed from the extracted RNA using standard Illumina libraries prep protocols. Sequencing was performed on Illumina HiSeq2000 platform in pair end. For ChIP-Seq, Cells (~1x10^7) for each sample were fixed in 1% formaldehyde for 10 min. The crosslink reaction was stopped by addition of glycine. Next, cells were washed 3 times with PBS. Then, cells were resuspended in lysis buffer and sonicated in Diagenode Bioruptor (Biosence) to fragments with length range of 200-500bp. Antibodies to H3K4me3 and H3K27me3 (Abcam) were added to supernatants to capture nucleosomes with these two histone modifications, respectively. The nucleosomal DNA was recovered by removal of histones at overnight incubation at 65℃ and purified using QIAquick PCR purification kit (QIAGEN, Maryland). The ChIP’ed DNA was used to construct libraries for sequencing using the ChIP-seq Sample Prep Kit (Illumina) according manufacturer’s instructions. The RNA sequencing libraries were constructed from the extracted RNA using standard Illumina libraries prep protocols. The ChIP’ed DNA was used to construct libraries for sequencing using the ChIP-seq Sample Prep Kit (Illumina) according manufacturer’s instructions.
identifiers.namespace: GEO
identifiers.id: GSM1697674