GSM1945878: siEZH1_H3K27me2; Homo sapiens; ChIP-Seq
For RNASeq 10ug total RNA was used, and mRNA purification was performed using oligo-dT magnic beads(NEBNext PolyA mRAN magnetic isolation module # E7490). mRNA was fragmented and reverse transcript into 200-400bp dscDNA for library construction(NEB # E7525 # E6111). For ChIPSeq chromatin was fragmented into single nucleosome by micrococcal nuclease, and H3K27me1(Millipore Cat. 17-643),H3K27me2(Active Motif Cat. 61435),H3K27me3(Millipore Cat. 17-622 ) ChIP grade antibody were used for ChIP. ChIPed DNA was eluted in TE buffer. Rubicon ThruPLEX DNA-seq 48D kit was used for RNASeq and ChIPSeq library construction. Brifely, about 100ng dscDNA was used as initiation for end repair and prepared for adaptor ligation. After adaptor ligation, PCR amplification with 15 cycles for each library and gel selection was used for library size selection and purification. For ChIPSeq library construction, 10ng-20ng ChIP eluted DNA was used as initiation and after adaptor ligation, PCR with 15-18 cycles was used to amplify library. Size selection of ChIPSeq library by electrophoresis gel.
GEO Accession
GSM1945878