This protocol outlines how to isolate, wash, and count nuclei suspensions for use with the Chromium Next GEM Single Cell Multiome ATAC + Gene Expression (GEX) protocol (CG000338). Cryopreserved primary cells (PBMCs) and cell lines (GM12878 cells; 3T3 cells) were used to develop this protocol. PBMCs were cryopreserved in IMDM + 40% FBS + 15% DMSO. Cell lines were cryopreserved in RPMI + 15% FBS + 5% DMSO. Optimization of some protocol steps (e.g. lysis time, centrifugation speed/time and filtration steps) may be needed based on cell type.
Caution: RNAse rich samples may need additional optimization. Please see: Can I use RNase-rich tissue samples for single-cell gene expression or Multiome assays?
FOR USE WITH:
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Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle, 16 rxns PN-1000283, includes:
- Chromium Next GEM Single Cell Multiome ATAC Kit A, 16 rxns PN-1000280
- Chromium Next GEM Single Cell Multiome Reagent Kit A, 16 rxns PN-1000232
- Library Construction Kit, 16 rxns PN-1000190
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Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle, 4 rxns PN-1000285, includes:
- Chromium Next GEM Single Cell Multiome ATAC Kit A, 4 rxns PN-1000281
- Chromium Next GEM Single Cell Multiome Reagent Kit A, 4 rxns PN-1000236
- Library Construction Kit B, 4 rxns PN-1000279
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Chromium Next GEM Chip J Single Cell Kit, 48 rxns PN-1000234
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Chromium Next GEM Chip J Single Cell Kit, 16 rxns PN-1000230
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Single Index Kit N Set A, 96 rxns PN-1000212
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Dual Index Kit TT Set A, 96 rxns PN-1000215