GSM5525259: CRISPRi-empty hPGCLC rep3; Homo sapiens; RNA-Seq
hPGCLC aggregates were dissociated with 0.05% Trypsin-EDTA for 10 min at 37°C. The dissociated cells were then stained with Integrin α6-Brilliant Violet 421 and EpCAM-AlexaFluor 488 conjugated antibodies, washed with FACS buffer (1% BSA in PBS) and resuspended in FACS buffer (1% BSA in PBS) with 7-AAD (BD PharMingen, 559925) as a viability dye. Cells were sorted using FACS into 350 uL of RLT Buffer (Qiagen). RNA was extracted using RNeasy micro kit (Qiagen). RNA-seq libraries were constructed by using Ovation Rapid Library Systems (Tecan, 0319-32) and quantified by a KAPA library quantification kit (Kapa Biosystems, kk4824).