GSM1364077: MJB144_RR_red-bisulphite_ESC_mouse_R1; Mus musculus; Bisulfite-Seq
Cells were resuspended in lysis buffer (100 mM Tris, pH 5.5, 5 mM EDTA, 200 mM NaCl, 0.2% SDS), supplemented with 400 µg/ml proteinase K (Invitrogen) and 200 µg/ml RNase A (Qiagen), and were incubated at 55°C overnight. DNA was extracted using phenol-chloroform and Phase Lock Gel Light tubes (5 Prime), was precipitated in 70% ethanol and resuspended in pure water. Samples of genomic DNA (12 x 1 µg) were digested with MspI (1 µL) in a digestion (30 µL total volume) overnight at 37 °C. To terminate the digestions, EDTA (3 µL, 0.5 M) was added. The DNA was subsequently worked up using the GeneJET PCR kit and eluted in EB (60 µL). The internal control DNA (1 µL, 0.6 ng/µL) was then added to all samples. All twelve MspI digested samples were subjected to an Illumina library preparation with the Illumina TruSeq kit using a 1/10 dilution of adapters. For the last AMPureXP purification from the TruSeq protocol the DNA was left to dry for 30 mins and eluted in water.
GEO Accession
GSM1364077