title: GSM4106212: Young uterus rep2; Mus musculus; MeDIP-Seq
library_construction_protocol: Genomic DNA was extracted from snap frozen tissue using Qiagen Allprep DNA/RNA mini kit (Qiagen, 80204), according to the manufacturers instructions. DNA libraries were generated using the NEBNext DNA library prep master mix set for Illumina (New England BioLabs E6040L), according to the manufacturerâ€™s instructions till the end repair stage, using 1.5Âµg of sonicated DNA. Unamplified DNA libraries were subjected to methylated DNA immunoprecipitation (MeDIP) to isolate fragments of methylated DNA. Briefly, library DNA was denatured at 99oC before incubation with anti-5mC antibody (Eurogentec BI-MECY-0100) for 2 hours at 4oC, followed by a further 2 hours with pre-blocked Dynabeads M-280 Sheep anti-mouse IgG (Invitrogen 112-01D). Methylated DNA was purified using the MinElute PCR purification kit (Qiagen 28104), amplified using the NEBNext DNA library prep master mix set, and size-selected for 200-800bp fragments using AMPure XP beads (Beckman Coulter A63881). Indexed libraries were pooled and sequenced on an Illumina HiSeq2500, using a 50bp paired-end protocol
attributes.tag: GEO Accession
attributes.value: GSM4106212