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Fig. 1 | Journal of Translational Medicine

Fig. 1

From: PBMC fixation and processing for Chromium single-cell RNA sequencing

Fig. 1

Experimental workflow of cell preparation and single-cell RNA-Seq. Donor PBMCs that were either live (a) or fixed (b, c) were analyzed their whole transcriptome at a single cell resolution. PBMCs in cold PBS (b, c) were fixed by adding 4 volumes of chilled methanol dropwise and then stored at − 20 or − 80 °C for up to 3 months. Right before scRNA-Seq, the PBMCs were resuspended in PBS (b) or SSC (saline sodium citrate, c). At the end of each step, the RNA quality was examined. b Fixed PBMC RNA was degraded during rehydration with PBS and could not generate sequencing library. c Fixed PBMCs resuspended in ×3 SSC preserved RNA integrity with RIN > 8.0 (confirmed in four different donors’ PBMCs) and were successfully implemented into 10× Chromium standard scRNA-Seq workflows as live PBMCs (a)

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