GSM8647358: Treg_CAR_P2Y3_donor31; Homo sapiens; RNA-Seq
Cells were immediately pelleted by centrifugation at 4°C and resuspended in Homogenization Buffer containing thioglycerol provided in the purification kit.  RNA was extracted with a Promega Maxwell RSC 16 using the Maxwell RSC simplyRNA Cells kit (Cat# AS1390) according to the manufacturer's instructions and RNA concentration was measured using a NanoDrop 8000.  RNA quality was assessed using an Agilent 4200 TapeStation and RINe values ranged from 9.9 to10. Total RNA (50 ng) was used the construction of libraries with the New England Biolabs NEBNext® Poly(A) mRNA Magnetic Isolation Module (Cat# 7490L) and Ultra II Directional RNA Library Prep Kit for Illumina (Cat# 7760L) according to the manufacturer's instructions.  Dual-indexed libraries were pooled to and sequenced at VANTAGE (Vanderbilt University) on an Illumina NovaSeq 6000 (S4 flow cell) to a depth of approximately 25 million paired-end 150 bp reads per library.