GSM9116227: GS_biol_rep_4; Homo sapiens; RNA-Seq
200 hematopoietic progenitor cells (CD34+, CD43+, CD33+ and DAPI -) derived from parental and mutant hiPSC were sorted at day 15 of EB differentiation directly in 8µl lysis buffer to 0.2% Triton X-100, and 2 U/μL RNase inhibitor (e.g., SUPERase•In, Thermo Fisher Scientific) Reverse Transcription (RT) and cDNA Amplification: The released RNA was reverse transcribed into complementary DNA (cDNA) using the Smart-seq2 protocol. Briefly, a template-switching oligo (TSO) (5'-AAGCAGTGGTATCAACGCAGAGTACATrGrG+G-3') was annealed to the RNA molecules, and reverse transcription was performed using SuperScript II reverse transcriptase (Invitrogen) at 42°C for 90 minutes, followed by 10 cycles of 50°C for 2 minutes and 42°C for 2 minutes. The cDNA was then purified using AMPure XP beads (Beckman Coulter) and amplified using KAPA HiFi HotStart ReadyMix (Kapa Biosystems) with 18 cycles of PCR amplification