GSM3081103: IgG ChIP control replicate 1; Homo sapiens; ChIP-Seq
ChIP assay was performed using SOLiD ChIP-Seq Kit (Applied Biosystems) according to manufacturer's guide. 1.2X10^7 SH-SY5Y cells were  suspended 500ul of PBS and subjected to chromatin cross-linking by 1% of formaldehyde for 10 minutes. The reaction was stopped by incubation with 57 ul of 1.25M glycine for 5 minutes and washing cells three times by PBS. Cells were lysed in 1ml of Lysis Buffer containing protease inhibitor and chromatin shearing was performed by sonication with Covaris S2 (5% duty cycle, for ten cycles, each 1 minutes, at 4oC intensity 2). Cell debris was removed by centrifugation at 20000g for 5 minutes. For immunoprecipitation, 50 ul supernatants were mixed 50 ul Dilution Buffer and incubated overnight at 4oC with 10 ul Dynabeads conjugated with anti-TCF4 antibodies K-12, N-16, 1G4 or control IgG. After washing with IP Buffers, beads were incubated with Proteinase K(370 ul/ml) at 55oC for one hour to reverse crosslinking and to elute DNA. ChIP-Seq library was prepared by end repairing ChIP DNA, ligating with P1 adaptor and barcode by T4 ligase and nick translating and amplification, each step followed by DNA purification by Agencourt AMPure XP beads. ChIP-seq, multiplex loading, single end sequencing, target 25 million raw tags per ChIP sample
GEO Accession
GSM3081103