GSM3521374: WT_DRIPseq_rep1; Saccharomyces cerevisiae; OTHER
Genomic DNA was extracted by spheroplasting and EtOH precipitation, excluding a RNase-A digestion. Purified DNA (50 μg) was digested by a cocktail of restriction enzymes (50 U HindIII, EcorI,  XbaI, SspI, BsrGI, NEB) in the presence or absence of 30 U of RNase-H (NEB) overnight at 37C. R-loops were enriched with the S9.6 antibody. DNA was end-repaired using a combination of T4 DNA polymerase, E. coli DNA Pol I large fragment (Klenow polymerase) and T4 polynucleotide kinase. A-tailing was performed with Klenow fragment (3' to 5' exo minus) and dATP. Illumina adapters were ligated and PCR amplification was performed with Illumina compatible primers for 13 cycles and library fragments were purified using 1:1 ratio of ampur XP beads.  Paired-end sequencing was performed on an Illumina NextSeq 500 with a read length of 75 bp.
GEO Accession
GSM3521374