ChIP-Atlas: SRX4665667

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: ChIP-seq was performed as previously described (Davies et al. Nature 2016) with several important modifications that increased ChIP stringency (noted here). Lysis was performed in 1% SDS lysis buffer (1% SDS, 10 mM EDTA, 50 mM Tris pH 8.0, 2x protease inhibitors). Sonication was performed in a Bioruptor Twin sonication bath at 4°C for three 5-minute periods of 30s on, 30s off at high power. Sonicated lysates were diluted 1:10 in IP wash buffer (100 mM Tris pH 7.5, 500 mM LiCl, 1% NP-40, 1% Na deoxycholate, 2x Protease Inhibitor, filtered) instead of dilution buffer for antibody incubation. This yielded roughly 1 ng of ChIP DNA per testis. Libraries were prepared using Illumina's TruSeq ChIP library preparation kit.