rabbit anti Spt5 (Glover-Cutter et al NSMB 15:71-78, 2008)
treatment
Xrn2 D235A expression was induced with 2ug/ml doxycycline for 24 hrs, Triptolide (10 mM) was added for 20 min.
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cross-linked chromatin was sonicated with a Bioruptor or Covaris Ultrasonicator 1-2 mg of cross-linked extract was immunoprecipitated, and prepared for illumina library construction by repair with Klenow, T4 DNA polymerase and T4 polynucleotide kinase, A-tailing with Klenow exo-, ligation to branched adaptors, SPRI bead purification of ~200bp fragments, and amplification (15-18 cycles) with Illumina Tru-seq primers using Phusion DNA polymerase.