GSM5032591: T107d_5month_Lead_M_Heart; Mus musculus; Bisulfite-Seq
Upon euthanasia at 5 months of age, heart were collected. Briefly, blood was removed by cardiac puncture (to be used in another study) and organs were perfused with PBS. Heart tissue was snap frozen in liquid nitrogen and stored at -80 deg C. Heart DNA extraction was performed using the AllPrep DNA/RNA/miRNA Universal Kit (Qiagen #80224). A total of 50 ng of genomic DNA was digested using restriction enzyme Mspl, and the digested DNA was then purified using phenol:chloroform extraction and ethanol precipitation in the presence of glycogen, before blunt-ending and phosphorylation. A single adenine nucleotide was next added to the 3′ end of the fragments in preparation for the ligation of the adapter duplex with a thymine overhang. The ligated fragments were cleaned, then processed for size selection on agarose gel. Selected fragments were treated with sodium bisulfite to convert unmethylated cytosines to uracils, which are then replaced with thymines during PCR amplification. These libraries were next cleaned up with AMPure XP beads (Product #A63880; Beckman Coulter), quantified using the Agilent TapeStation genomic DNA kit (Catalog #G2991AA; Agilent) and Qubit High Sensitivity dsDNA (Catalog #Q32850; Invitrogen).
GEO
GSM5032591