GSM6281277: A005_9_Input; Mus musculus; ChIP-Seq
Small pieces of frozen liver tissue from two ad libitum-fed male C57Bl/6 mice were pulverized in liquid nitrogen, and cross-linked with 1% formaldehyde for 10 min. After quenching with 2.5 M glycine, nuclear extracts were prepared. Chromatin fragmentation was performed by sonication for 20 cycles of 30 s at the highest level. Proteins were immunoprecipitated using anti-BHLHE40 (Novus Bio NB100-1800) antibody and anti-ChREBP (Novus Bio NB400-135) antibody. Antibodies were precipitated, crosslinked and DNA was isolated using phenol/chloroform/isoamyl alcohol. As control sample input DNA was used. ChIPseq was performed with an average depth of 20 million reads per sample, 75 bp single end reads (75bp cycle kit), on the Illumina NextSeq500.