alias: GSM2988432
title: GSM2988432: Input; Mus musculus; ChIP-Seq
library_construction_protocol: ChIP assays were performed with EZ-ChIP™ Chromatin Immunoprecipitation Kit (Millipore, #17-371). 5 × 105 sorted mTECs of wild-type mice and 2μg antibodies were used for each immunoprecipitation. ChIPed DNA was amplified by SeqPlexTM Enhanced DNA Amplification Kit (Sigma Aldrich, SEQXE) according to manufacturer's protocol and sequenced Libraries were prepared according to Illumina's instructions. 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 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, library fragments were band isolated from an agarose gel. Libraries were sequenced on the Illumina HiSeq™2500 following the manufacturer's protocols.
identifiers.namespace: GEO
identifiers.id: GSM2988432