GSM8513516: CFs, Control, rep2; Mus musculus; RNA-Seq
Total RNA was extracted using Trizol reagent and assessed for quantity and purity with a Bioanalyzer 2100. High-quality RNA (RIN > 7.0) was used for library construction. mRNA was purified from 5 µg of total RNA using Dynabeads Oligo (dT) and fragmented with the Magnesium RNA Fragmentation Module. RNA fragments were reverse-transcribed into cDNA using SuperScript™ II Reverse Transcriptase and converted into U-labeled second-stranded DNAs with E. coli DNA polymerase I and RNase H. These were ligated to dual-index adapters and size-selected with AMPureXP beads. After UDG treatment, the products were PCR-amplified and sequenced with 2×150 bp paired-end reads on an Illumina NovaSeq 6000.