ChIP-Atlas: SRX663448

Antigen

Cell type Class: Gonad
Cell type: Testis
MeSH Description: The male gonad containing two functional parts: the SEMINIFEROUS TUBULES for the production and transport of male germ cells (SPERMATOGENESIS) and the interstitial compartment containing LEYDIG CELLS that produce ANDROGENS.

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: For H3K4me3 CHiP-Seq and DMC1 SSDS protocols, testicular samples were obtained frozen and were directly thawed in 1% paraformaldehyde and gently dissociated. Genomic DNA for whole genome sequencing was extracted from the testicular samples before fixation with the DNeasy Blood and Tissue kit (Qiagen). DMC1 and H3K4Me3 ChIP were performed as described previously (Smagulova et al., 2011; Khil et al., 2012; Brick et al., 2012) with minor modifications. Sequencing libraries for anti-H3K4Me3 samples were prepared according to manufacturer’s protocol (Illumina) and anti-DMC1 libraries were prepared following the method described in (Khil et al., 2012). For genomic DNA sequencing, DNA was sheared using a Covaris S series (Covaris, MS, USA) focused-ultrasonicator. Standard sequencing library construction was done using enzymes from New England Biolabs according to the protocol provided by Illumina. The PCR amplified library was size selected on a 2% agarose gel to extract the 300 to 500 bp band.