ChIP-Atlas: SRX3630649

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 DMC1 SSDS, 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 ChIP was performed as described previously (Smagulova et al., 2011; Khil et al., 2012; Brick et al., 2012) with minor modifications. For H3K4me3 ChIP-Seq, nuclei were isolated from cells using hypotonic lysis. Chromatin was fragmented by MNase digestion. After high speed spin soluble chromatin was removed and used for ChIP. Sequencing libraries for anti-DMC1 were prepared following the method described in (Khil et al., 2012). For H3K4me3 ChIP-Seq, libraries were prepared for sequencing using Bio Scientific's NEXTflex ChIP-Seq Kit (protocol version V11.11) without size selection. Amplification of the libraries was done with 20 µL ligation product, 16 ul water, 12 µL NEXTflex ChIP PCR master mix, 2 µL NEXTflex ChIP primer mix and 14-18 cycles of PCR. ChIP-Seq, Single Stranded DNA Sequencing (SSDS)