This protocol outlines how to isolate, wash, and count single nuclei from fresh, cryopreserved, and flash frozen embryonic mouse brain tissue samples for use with the Chromium Single Cell Multiome ATAC + Gene Expression solution. Tissue triturated into a nearly single cell suspension and subsequently frozen in media containing 10% DMSO produced metrics comparable to fresh tissue. High quality data can also be obtained using flash frozen tissue. Optimization of some protocol steps (e.g. homogenization, lysis reagent/time, centrifugation speed/time and filtration steps) may be needed when working with mouse brain tissues from different sources.
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