GSM4144629: BisSeq_infect_3T3_rep1; Mus musculus; Murine leukemia virus; OTHER
To prepare virus for bisulfite sequencing, viral particles were first concentrated by sedimenting filtered media from infected cells through 2-mL cushions of 20% sucrose and viral RNAs isolated using Trizol (Invitrogen. For infected cellular samples, total RNA was extracted in Trizol. Cellular and viral RNAs were DNase treated. Additionally, cellular samples were subjected to RiboMinus (Thermo) treatment to deplete ribosomal RNAs. The Zymogen EZ RNA Methylation kit protocol was followed to convert unmethylated cytosines to uracil. Recovered RNA was used to generate sequencing libraries with the TruSeq Stranded mRNA kit (Illumina). cDNA libraries were amplified by 12 cycles of PCR, loaded onto a HiSeq 2000 Illumina sequencer, and sequenced for 100 bases at The University of Chicago Genomics Facility. RNA bisulfite sequencing
GEO Accession
GSM4144629