SRX2175445 GSM2320047 SRP090123 SRS1702733 GSM2320047 MNase-Seq GENOMIC MNase Mono-nucleosomal DNA was isolated from total meiotic populations of 10-week old B6 mouse male testes (1.3 x 106 cells) by MNase digestion of native chromatin. Final mono-nucleosomal DNA was purified using phenol/chloroform extraction followed by ethanol precipitation, run on 2 % agarose gel and isolated using Qiagen gel extract kit. Gel-purified mono-nucleosomal DNA was then used as a template for sequencing library construction. Library was prepared according to Illumina's instructions. Briefly, DNA was end-repaired using a combination of T4 DNA polymerase, E. coli DNA Pol I large fragment (Klenow polymerase) and T4 polynucleotide kinase. The blunt, phosphorylated ends were treated with Klenow fragment (32 to 52 exo minus) and dATP to yield a protruding 3- 'A' base for ligation of Illumina's adapters which have a single 'T' base overhang at the 3’ end. After adapter ligation DNA was PCR amplified with Illumina primers for 15 cycles and library fragments of ~250 bp (insert plus adaptor and PCR primer sequences) were band isolated from an agarose gel. The purified DNA was captured on an Illumina flow cell for cluster generation. The library was sequenced on the Genome Analyzer following the manufacturer's protocols. Illumina Genome Analyzer IIx gds 302320047 GEO Accession GSM2320047