GSM4453029
GSM4453029: C2C12, input sample; Mus musculus; ChIP-Seq
Chromatin immunoprecipitation were performed as in Buschbeck et al., 2009. 10 ng of enriched DNA was used for library generation and parallel sequencing on Illumina genome Analyzer at the CRG Genomics Unit (CRG Core Facilities). DNA quality was controled by High Sensitivity Qubit. From 10 ng of DNA, the sequencing was performed on single read (SR), with an average length of reads of 50 bp, 30 000 reads per sample, on an Illumina HiSeq2500 sequencer. 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 (32 to 52 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 15 cycles and library fragments of ~250 bp (insert plus adaptor and PCR primer sequences) were band isolated from an agarose gel. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the Genome Analyzer following the manufacturer's protocols.
GEO
GSM4453029
GEO Accession
GSM4453029