GSM4585144: timecourse-23C-30min; Saccharomyces cerevisiae; OTHER
Micro-C products were extracted as the following steps: 1) Digest crosslinked chromatin by MNase; 2) Repair fragment ends with biotin-dNTP by DNA Polymerase I, Large (Klenow) Fragment ; 3) Proximal ligation by T4 DNA ligase and remove unligated ends by Exonuclease III; 4) Purify ligated dinucleosomal DNA by the standard Phenol:Chloroform:Isoamyl Alcohol (25:24:1) and ethanol precipitation procedure. Purified DNA with biotin-dNTPs was captured by Dynabeads® MyOne™ Streptavidin C1. Standard library preparation protocol including end-repair, A-tailing, and adapter ligation was performed on beads with the individual enzymes purchased from Lucigen or the NEBnext UltraII kit. Sequencing library was amplified by Kapa HiFi PCR enzyme or NEB Q5 enzyme with the lowest possible cycles to reduce PCR duplicates. Sequencing library was sequenced paired-end 50x50 on the Illumina HiSeq 4000 sequencer.