title: GSM5066733: GFP overexpressing isolated muscle satellite cells RNA-seq Rep2; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted from Gfp or Fos-expressing primary muscle progenitor cells using TRIzol followed by RNeasy Mini Qiagen extraction kit according to manufacturer's protocol. In-situ Hi-C libraries were generated using the HindIII restriction enzyme (PMID: 22652625). Briefly, ~25 million cells were crosslinked with 1% formaldehyde for 10 minutes at room temperature. Then, the chromatin was extracted, digested with HindIII, end-labelled with biotin-14-dCTP, followed by  in-situ ligation (EL0011, Thermo Fisher). After DNA extraction, biotin was removed from unligated ends, and the sample was sheared using a Covaris S220 instrument (100-500bp range) as previously described (PMID: 22652625). After A-tailing, biotin pull-down, and adapter ligation, we performed paired-end sequencing on Illumina's HiSeq 2000 instrument. RNA-seq libraries were generated using the Illumina TruSeq (version 2) kit and sequenced on the Illumina HiSeq2500 instrument. RNA-seq libraries were prepared with polyA enrichment.