GSM1937508: SHP ChIP-seq with vehicle treatment; Mus musculus; ChIP-Seq
Liver tissues were chopped by a razor blade and cross-linked in 1% (vol/vol) formaldehyde for 10 min at room temperature with rotation. Then, glycine to a concentration of 0.125 M was added to quench the cross-linking reaction. The chopped liver tissues were washed with ice-cold PBS and homogenized in hypotonic solution. Nuclei were isolated and chromatin DNA was sheared by sonication (Fisher Scientific, model 100). Four ug of SHP antibody (sc-30169, Santa Cruz) was added to the sheared chromatin and 10% of the sheared chromatin was saved for input. After incubation overnight at 4oC, protein G agarose beads were added and the incubation was continued for 1 h at 4°C with rotation. Then, the precipitated protein G agarose beads-antibody complex was washed by sodium- and lithium-chloride buffers. The input and the precipitated complexes were treated with RNase A and Proteinase K, and incubated at 65°C overnight to reverse crosslinks. DNA was isolated by phenol-chloroform extraction. The ChIPseq libraries were prepared with Illumina's 'TruSeq DNaseq Sample Prep kit' with two modifications: adaptors were diluted 1:20 and adaptored DNAs were amplified for 15 cycles. The libraries were pooled in equimolar concentration and the pool was quantitated by qPCR and sequenced on one lane for 101 cyclesusing a TruSeq SBS sequencing kit version 3 and analyzed with Casava 1.8.2
GEO Accession
GSM1937508