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What 10x Genomics workflows can be run on the Chromium Connect?

Question: What 10x Genomics workflows can be run on the Chromium Connect?

Answer: The document LIT000154_Chromium-Connect-Kit-Selection-Guide outlines the Supported workflows for:

  • Continuous or modular cells-to-libraries Single Cell 3’ and 5’ gene expression workflows from cell or nuclei suspensions
  • Library construction or 5’ V(D)J amplification workflows (TCR/BCR profiling) from purified cDNA
  • 5’ Feature Barcode library construction from 5’ Feature Barcode reaction amplified DNA
  • Library construction from Single Cell gene expression Flex (Fixed RNA) pre-amplification sample

Unsupported library construction workflows may be run as described below, at users’ own risk. Unsupported automated workflows have not been verified nor validated for the starting samples of interest but provide the operator with the means to process their manual samples using kits and automated workflows that were validated for other sample types.  These unsupported workflows include the following (below Disclaimer):

 

Disclaimer

a.     The use of 10x Genomics products in practicing the methods set forth herein has not been validated by 10x Genomics, and such non-validated use is not covered by 10x genomics standard warranty, and 10x Genomics hereby disclaims any and all warranties for such use.

b.      Nothing in this article should be construed as altering, waiving or amending in any manner 10x Genomics terms and conditions of sale for the Chromium Connect instrument, consumables or software, including without limitation such terms and conditions relating to certain use restrictions, limited license, warranty and limitation of liability, and nothing in this article shall be deemed to be Documentation, as that term is set forth in such terms and conditions of sale. Nothing in this article shall be construed as any representation by 10x Genomics that it currently or will at any time in the future offer or in any way support any application set forth herein.

 

  • 3’ and 5’ Single Cell HT gene expression library construction
    • Starting material should be 20 μl of purified cDNA sample (manually processed from GEMs from a Chromium X, see User Guides CG000416 or CG000423) put into the ODTC full-skirted plate column 7.  
    • Automated Kit materials needed are the same as for Single Cell 3’ or 5’ Gene Expression Purified cDNA listed on p3 of LIT000154:
        • Automated Library Construction Kit
            • 4 rxns: Product code 1000429
            • 24 rxns: Product code 1000428
        • Dual Index Kit TT Set A
            • 96 rxns: Product code 1000215
      • Follow the instructions for Automated Gene Expression Library Construction outlined in CG000474 and on-screen select Library Construction > Single Cell GEX Kit on the Chromium Connect.
      • Follow the Sample Index PCR cycle number selection guidelines in CG000416 (manual Single Cell 3’ HT Gene Expression User Guide) or CG000423 (manual Single Cell 5’ HT Gene Expression v2 User Guide), based on cDNA input mass. Enter this Sample Index PCR cycle number selection into the Chromium Connect Software.
      • At completion of the run, libraries will be in tube strip 4.
  • Multiome gene expression library construction
    Note: This method is to be used on the Chromium Connect to construct gene expression libraries from pre-amplified transposed DNA/cDNA obtained using the 10x Genomics Chromium Next GEM Single Cell Multiome ATAC + Gene Expression manual workflow.     Multiome ATAC library construction is not enabled on the Chromium Connect.
    • Starting material should be 10 μl of the amplified, purified cDNA sample (manually processed through the Chromium Next GEM Single Cell Multiome ATAC + Gene Expression protocol CG000338) put into the ODTC full-skirted plate column 7. 
    • Automated Kit materials needed are the same as for Single Cell 3’ or 5’ Gene Expression Purified cDNA listed on p3 of LIT000154:
        • Automated Library Construction Kit
            • 4 rxns: Product code 1000429
            • 24 rxns: Product code 1000428
        • Dual Index Kit TT Set A
            • 96 rxns: Product code 1000215
      • Follow the instructions for Automated Gene Expression Library Construction outlined in CG000474 and on-screen select Library Construction > Single Cell GEX Kit on the Chromium Connect.
      • Follow the Sample Index PCR cycle number selection guidelines in CG000338 (manual Chromium Next GEM Single Cell Multiome ATAC + Gene Expression User Guide), based on cDNA input mass and high/low RNA content. Enter this Sample Index PCR cycle number selection into the Chromium Connect software.
      • At completion of the run, libraries will be in tube strip 4.

Products: Chromium Connect, Single Cell Gene Expression, Single Cell Immune Profiling, Single Cell Multiome ATAC+GEX