GSM9084847: bf23, Remission_pts4; Homo sapiens; RNA-Seq
MNCs were washed thrice with 1X Phosphate-Buffered Saline (PBS, free of calcium and magnesium) containing0.04% weight/volume FBS (Gibco, Cat# 70011-044). Thereafter MNCs were counted by the Burker chamber. CD138+ BM cells were prepared using the EasySep CD138 Positive selection kit II (STEMCELL Technologies, Cat#17887A), after erythrocyte lysis (Cytognos, Cat# CYT-BL), following manufacturer's instructions. The Chromium Next GEM Single Cell 5' v2 protocol (CG000330 Rev E) was used to calculate the volume of cells to load for each sample, with the aim of recovering 9,000 cells. The Chromium Next GEM Chip K (10X Genomics; PN-2000286) with Master Mix from Chromium Next GEM Single Cell 5' Reagent Kits v2 (Dual Index) (10X Genomics; PN-1000263) and Chromium Single Cell Human TCR Amplification Kit, (10X Genomics; PN-1000252) were employed to generate Single-cell 5′ RNA-seq libraries. After the GEM was generated and the barcoded full-length cDNA amplificated, the SPRIselect Reagent (Cat# B23318) was used to purify cDNA. The latter was then quantified by Picogreen dsDNA assay kit (Cat# P7589). To assess quality, the Agilent 4150 TapeStation machine was employed by using High Sensitivity D5000 ScreenTape (Agilent, Cat# 5067-5592) with High Sensitivity D5000 Reagents (Agilent, Cat# 5067-5593).
GEO
GSM9084847_r1