RNA-Seq of small cell lung cancer cell line
Total RNA was isolated from fresh cells using TRIzol Reagent (Invitrogen, 15596026). 500ng total RNA spiked in with Sequins (Garvan Institute) and SIRV-Set 4 (Lexogen, 141.01) was used to prepare libraries with the TruSeq Stranded Library Prep Kit (Illumina). Superscript III (Invitrogen, 18080093) was used in place of Superscript II for reverse transcription with RT temperature modified to 50C. Fragmentation was performed at 94C for 4 minutes, and final amplification with 15 PCR cycles. The libraries were sequenced in 100PE on the NextSeq 500.