Diseased whole blood was obtained by 10x Genomics from Sanguine Biosciences.
Peripheral blood mononuclear cells (PBMCs) from a diseased Primary Myelofibrosis, JAK2+ donor were isolated following CG000392 Rev B. Fresh PBMCs were directly processed and gene expression libraries were generated as outlined in the Chromium GEM-X Single Cell 3' Reagent Kits v4 User Guide (CG000731) or fixed following the demonstrated protocol Cell Fixation Protocol for GEM-X Single Cell 3' & 5' Assays (CG000776). Fixed PBMCs were processed at two timepoints, (1) immediately post fixation, and (2) after one storage for 1 month at -80°C.
Libraries were sequenced on a NovaSeq 6000 with the following sequencing configuration:
- 28 cycles Read 1
- 10 cycles i7
- 10 cycles i5
- 90 cycles Read 2
Fixation offers opportunities to streamline processes and enhance consistency in your workflows. Our demonstrated protocol is specifically designed and optimized for GEM-X 3’ and 5’. It is important to note that while there are differences in data for fixed samples compared to fresh samples, Gene Expression and V(D)J information is maintained in fixed samples.
See companion datasets