GSM4211237: sWGA-MCF-7-Input; Homo sapiens; ChIP-Seq
MCF-7 and MCF-7/ADR cells were maintained in RPMI-1640 medium with 10% fetal calf serum. Approximately 2 x 10e7 cells were used for each COGC-seq or sWGA-seq assay. O-GlcNAz chromatin DNAs enriched by Streptavidin-magnetic beads were purified with the Qiagen PCR purification kit. For sWGA-seq, O-GlcNAc protein bound chromatin DNAs were enriched by sWGA-agarose beads. For RNA-seq, total RNA was extracted from the MCF-7 and MCF-7/ADR cells using Trizol reagent (Invitrogen). RNA quality was checked by Agilent 2100 Bioanalyzer (Agilent Technologies, USA). RNA libraries were prepared for sequencing using standard BGI protocols. DNA-end repair, 3'-dA overhang and ligation of methylated sequencing adaptor. PCR amplification and size selection (usually 100-300bp, including adaptor sequence. Qualified library for sequencing. RNA-seq libraries were constructed using SMARTer cDNA library construction kit (Clontech, Takara Bio, CA, USA).
GEO Accession
GSM4211237