title: GSM4332476: 5hMeDIP_HellscKO_2; Mus musculus; MeDIP-Seq
library_construction_protocol: For DMC1-SSDS experiment : Cell suspension was prepared by filtering homogenized tissue, then lysed. Lysates were sonicated and chromatin complexes were isolated with appropriate antibody. For 5hMeDIP-seq : genomic DNA was extracted by phenol/chloroform extraction  from a population of  ~95% pure leptotene/zygotene (L/Z) spermatocytes, obtained from synchronized FAC-sorted testes. For DMC1 experiment, standard library construction was performed with the Illumina Trueseq protocol (Illumina, IP-202-9001DOC) with an additional step of kinetic enrichment as described in Khil et al. 2012. For 5hMeDIP-seq, Illumina adaptors were added to sonicated gDNA with NEB Next Ultra library preparation kit (NEB7370S), without the final PCR step. hMeDIP was then performed with Active Motif hMeDIP kit (AM, 55010) according to the manufacturers' manual. Enriched fragments were then amplified by PCR, as recommended by the NEB Next Ultra library preparation kit. Single Stranded DNA Sequencing (SSDS) method described in Khil et al. 2012 for DMC1 ChIPseq ; 5hMeDIP for 5hMeDIP-seq experiment
identifiers.namespace: GEO
identifiers.id: GSM4332476