GSM7697115: Sample_7; Homo sapiens; RNA-Seq
200 CD34+ lineage negative cells were isolated by FACS and sorted directly into 8 uL of lysis buffer (0.2% Triton X-100, Sigma) containing oligo-dT primers (2.5 uM, Integrated DNA Technologies UK Ltd, Cat #51-01-15-01), dNTP mix (2.5 uM, Life Technologies, Cat#19155) and RNase inhibitor (10 U/ul, Takara (Clontech), Cat# 2313A) aliquoted into 96 well PCR plate (Thermo Fisher segmented semi-skirted Cat #AB-0900) on ice. Cell lysis, RT and PCR amplification (20 cycles) was performed using the Smart-Seq 2 kit (SMARTScribe, Takara(Clontech) #639537). Libraries were pooled and sequenced on a on the NextSeq 500 platform (Illumina) using a NextSeq 500/550 High Output v2.5 (75 cycle) sequencing kit (Illumina 20024906), generating 75-bp single-end reads.