GSM6248777: 3s170_High; Homo sapiens; RNA-Seq
RNA was not isolated from cells. Instead, sequencing libraries were prepared directly from cell lysates. Indexed, directional sequencing libraries were prepared directly from intact cells using the SMART-Seq Stranded Kit (Takara Bio, Inc.) according to the manufacturer's standard protocol. Briefly, 300 cells were lysed and followed by random-primed first-strand cDNA synthesis, tailing, and template switching with reverse transcriptase (SMARTScribe RT). Illumina barcoded adapters were incorporated by PCR with SeqAmp DNA Polymerase and rRNA-derived cDNA was selectively depleted. The remaining mRNA-derived cDNA fragments were PCR-enriched and the resulting libraries purified with AMPure XP beads (Beckman Coulter, Indianapolis, IN).