title: GSM2052338: SB-431542 1 SMAD2 ChIP; Mus musculus; ChIP-Seq
library_construction_protocol: Samples for ChIP-seq with the corresponding inputs were prepared by crosslinking in 1% formaldehyde for 10 minutes at RT, cytoplasmic lysis, sonication of nuclei to shear the DNA to 100-500 bp, followed by incubation with anti-SMAD2 antibody overnight. pre-blocked protein A dynabeads (invitrogen) were added for 4 hours, followed by washing, DNA elution, reversal of crosslinks and proteinase K digestion. Resulting enriched ChIP DNA was cleaned up using Qiagen PCR purification kit. Corresponding input chromatin was processed alongside at the same time. Following end repair, poly-A-tailing and adapter ligation, Illumina TruSeq ChIP sample preparation kits were used to generate the libraries. The Illumina kit Phusion enzyme was replaced by Kapa HiFi HotStart ready mix (Kapa Biosystems, Cape Town, South Africa).  The PCR was run before gel isolation using the Invitrogen SizeSelect E-gel system (SizeSelect gel protocol, Thermo Fisher Scientific).  Post PCR we used AMPure XP beads (AMPure bead protocol, Beckman Coulter, Inc.) at a 1:1 ratio to maintain size integrity. Samples were multiplexed and 51-bp single end reads were generated on an Illumina HiSeq 2500.
attributes.tag: GEO Accession
attributes.value: GSM2052338