title: GSM2829751: insitu3Cseq_HindIII_Scer_Async_12 _2hr_37C_elegansspike_rep1; Saccharomyces cerevisiae; Caenorhabditis elegans; OTHER
library_construction_protocol: 3C-seq: Cells were fixed in 3% FA. Cells were then spheroblasted with zymolase and Nuclei were extracted with douncing. Nuclei were counted after staining with Crystal Violet. At this point if a spike-in was used (ycs4-2 experiments), 10% of N2 embryonis C.elegans nuclei were added. Nuclei were permeabilised and then the restriction enzyme HindIII was used to digest the genome. Digested genomic DNA was subjected to proximity ligation using a T4 DNA Ligase in dilute condtions. DNA was then purifed using Qiagen PCR purifaction columns and DNA sonicated down to 200bp-1kb size with a Diagenode Bioruptor. DNA was ligated to Illumina or home-made multiplexed adapters and amplified by PCR. For 3C-seq library DNA between 400-1Kb was gel purified. For ChIP Library DNA between 250-500 bp in size was gel purified.
identifiers.namespace: GEO
identifiers.id: GSM2829751