Hap1 NELFA-degron (Sheridan and Bentley Biotechniques 60:69-74, 2016)
cell line/genotype
Hap1 NELFA-degron (Sheridan and Bentley Biotechniques 60:69-74, 2016)
spike in
NA
growth conditions
IMDM 10% FCS, 10mM trimethoprim (note NELFA degron not induced) NaCl 350mM 5 min.
antibody
rabbit anti pan CTD (Schroeder eta l Genes Dev. 14:2435, 2000)
experiment
Chromatin IP against RNA pol II
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
cross-linked Chromatin was sonicated with a Bioruptor 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, gel purification of ~200bp fragments, and amplification (18cycles) with Illumina Tru-seq primers using Phusion DNA polymerase.