title: GSM1138594: BMI1_CST_Scr_ChIPSeq; Homo sapiens; ChIP-Seq
library_construction_protocol: Lysates were clarified from sonicated nuclei and protein-DNA complexes were isolated with antibody. ChIP was performed using HighCell ChIP kit following manufacturers protocol (Diagenode) ChIP-Seq libraries were prepared using standard Illumina protocol. ChIP-enriched DNA samples (10-20 ng) were converted to blunt-ended fragments using a combination of T4 DNA polymerase, E.coli DNA polymerase I large fragment (Klenow polymerase) and T4 polynuleotide kinase (New England BioLabs, NEB). A single A-base was added to fragment ends by Klenow fragment (3' to 5' exo minus; NEB) followed by ligation of Illumina adaptors (Quick ligase, NEB). The adaptor-modified DNA fragments were enriched by PCR using the Illumina PE1.0/PE2.0 primers and Phusion DNA polymerase (NEB). PCR products were size selected using 3% NuSieve agarose gels (Lonza) followed by gel extraction using QIAEX II reagents (QIAGEN). Libraries were quantified using the Bioanalyzer 2100 (Agilent) and each sample was sequenced in a single lane on the Illumina HiSeq 2000 (40 nucleotide read length).
attributes.tag: GEO Accession
attributes.value: GSM1138594