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Nuclei Isolation from Complex Tissues for Single Cell Multiome ATAC + Gene Expression Sequencing

  • CG000375_DemonstratedProtocol_NucleiIsolationComplexSample_ATAC_GEX_Sequencing_Rev C.pdf

    Demonstrated Protocol, CG000375

    CG000375_DemonstratedProtocol_NucleiIsolationComplexSample_ATAC_GEX_Sequencing_Rev C.pdf

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This protocol outlines how to isolate, wash, and count nuclei suspensions from complex tissues for use with the Chromium Next GEM Single Cell Multiome ATAC + Gene Expression (GEX) protocol (CG000338). Fresh frozen human malignant lymphoma, glioblastoma, and normal brain tissue were used to develop this protocol. 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?

Important
This protocol is only compatible with the Chromium Single Cell Multiome ATAC + Gene Expression reagent kits. Do not use with standalone Single Cell ATAC or SC3'v3.1 assays.

FOR USE WITH:

  • 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
  • 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
  • Chromium Next GEM Chip J Single Cell Kit, 48 rxns PN-1000234

  • Chromium Next GEM Chip J Single Cell Kit, 16 rxns PN-1000230

  • Single Index Kit N Set A, 96 rxns PN-1000212

  • Dual Index Kit TT Set A, 96 rxns PN-1000215

Document Type
Demonstrated Protocol

Last Modified
July 12, 2022