GSM4063760: WT_4; Mus musculus; ChIP-Seq
10 million activated cells were crosslinked with 1% formaldehyde. Cells were lysed according to the protocol of truChIP Chromatin Shearing Kit with Formaldehyde (Covaris). Cell lysates were sonicated and precleared lysate was incubated with 2 g of anti-AID antibody at 4oC overnight. Immunocomplexes were recovered by incubation with Salmon Sperm DNA/Protein A Agarose Slurry, eluted, then de-crosslinked with NaCl and RNase A at 65oC for 4-6 hours, and treated with Tris pH 6.5, EDTA and proteinase K for 1 hour at 45oC. DNA was recovered by phenol-chloroform extraction and ethanol precipitation. See manuscript for more details. Immunoprecipitated DNA was quantified by PicoGreen and the size was evaluated by Agilent BioAnalyzer. Illumina libraries were prepared using the KAPA HTP Library Preparation Kit (Kapa Biosystems KK8234) according to the manufacturer's instructions with 0 (undetectable)-4.6 ng input DNA and 8-12 cycles of PCR. Barcoded libraries were run on the Hiseq 2500 in Rapid mode and the HiSeq 4000 in 50bp/50bp paired end runs, using the HiSeq Rapid SBS Kit v2 and HiSeq 3000/4000 SBS Kit, respectively (Illumina). An average of 35 million paired reads were generated per sample.
GEO
GSM4063760
GEO Accession
GSM4063760