GSM1948530: siSPOP_RNASeq; Homo sapiens; RNA-Seq
10ug total RNA was used for RNASeq library construction. mRNA was purified by oligo-dT magnetic beads and fragmented into 200-400bp by NEBNext RNA processing kit(NEB E7525,E7490,E6111), and then reverse transcript to dscDNA. For ChIP assay, DNA and protein were cross-link and digested into single nucleosome by using micrococcal nuclease, and then H3K36me3(abcam #ab9050) ChIP grade antibody was used for IP enrichment. ChIP DNA was reverse cross-link at 65°C overnight and  eluted by TE buffer. We used Rubicon ThruPLEX DNA-seq(Rubicon #R400406) for library construction.For RNASeq library construction, about 100ng total fragmented dscDNA as initiation for end repair,adaptor ligation and PCR amplification with 15 cycles.After amplification gel selection was used to purify RNASeq library. For ChIPSeq library construction, about 20ng ChIPed DNA as initiation. The PCR amplification cycles for ChIPSeq was 18 and gel selection for library purification.
GEO Accession
GSM1948530