GSM4337022: HBV infected hepatocytes rep1; Homo sapiens; Mus musculus; RNA-Seq
RNA from four HBV-infected and three uninfected liver chimeric mice were collected using standard procedures and the RNeasy Qiagen kit for library production and strand-specific RNA-sequencing at the Beijing Genomics Institute (BGI, Hong Kong). RNA purity was assessed with a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific) and RNA integrity was determined with a 2100 Bioanalyzer Instrument (Agilent Technologies). Poly (A) mRNA was purified from total RNA using magnetic beads with oligo (dT) (New England BioLabs, Hitchin, UK). The mRNA was then broken into short fragments using divalent cations at an elevated temperature. The cleaved RNA fragments are copied into first strand cDNA using reverse transcriptase and random primers. This is followed by second strand cDNA synthesis using DNA Polymerase I and RNaseH (New England BioLabs, Hitchin, UK). These cDNA fragments then have the addition of a single 'A' base and subsequent ligation of the adapter. The products are purified and enriched with PCR amplification. The library is then circularized as previously described (Drmanac, R et al. 2010). From single stranded circles, DNA nanoballs are made and loaded onto patterned nanoarrays (Drmanac, R et al. 2010). The nanoarrays are then subjected to combinatorial probe-anchor synthesis (cPAS) based sequencing on the BGISEQ-500 (Fehlmann, T. et al, 2016; Huang, J. et al, 2017; Mak, S. S. T. et al, 2017), employing the PE100 mode to produce raw paired‐end reads of 100 bp.
GEO Accession
GSM4337022