GSM1841304: RS411_Dex1h_DHS; Homo sapiens; DNase-Hypersensitivity
Histone modification ChIP and DHS were performed on MNase-digested and DNase-digested native chromatin, respectively.  Transcription factor ChIP was performed on sonicated nuclei isolated from cells treated with EGS and formaldehyde cross-linking agents. Libraries were prepared according to Illumina's instructions accompanying the DNA Sample Kit (Part# 0801-0303). Briefly, DNA was end-repaired using a combination of T4 DNA polymerase, E. coli DNA Pol I large fragment (Klenow polymerase) and T4 polynucleotide kinase. The blunt, phosphorylated ends were treated with Klenow fragment (3' to 5' exo minus) and dATP to yield a protruding 3- 'A' base for ligation of Illumina's adapters which have a single 'T' base overhang at the 3’ end. After adapter ligation DNA was PCR amplified with Illumina primers for 7-9 cycles and library fragments of ~270 bp for histone modification libraries, ~75 bp for DHS libraries, and ~320 bp for transcription factor libraries (insert plus adaptor and PCR primer sequences) were size selected using PippinPrep (Sage Science) 2% agarose gels. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the HiSeq2000 following the manufacturer's protocols.
GEO Accession
GSM1841304