GSM3358197: Ngn3 heterozygous islet progenitors; Mus musculus; Bisulfite-Seq
Endocrine cells were  were enriched by flow sorting for Ngn3eGFP expressing cells. Cells were > 95% viable. Cells were directly lysed for DNA preparation. Genomic DNA was prepared using a Qiagen genomic DNA isolation kit. Transposomes were assembled with Tn5 enzyme purified in house using a plasmid construct kindly provided by the Sandberg lab and according to protocols previously described (Picelli et al., 2014). Tagmentation, oligonucleotide replacement and gap repair were carried out on 75ng of genomic DNA. Bisulfite conversion of DNA was performed using the EZ DNA Methylation Lightning kit (Zymo cat # D5030) according to the manufacturers recommendations. After desulphonation and purification of sodium bisulfite treated libraries, samples were amplified for 12 cycles with the Kapa HiFi HotStart Uracil+ Ready Mix according to the manufacturers instructions. WGBS libraries were sequenced on the Illumina Novaseq (paired end 150bp read lengths), obtaining approximately 400 million reads per library.
GEO
GSM3358197