Aug 16, 2023

Public workspaceU54 SCENT 10x Genomic Single-Cell Multiome

DOI
dx.doi.org/10.17504/protocols.io.81wgbxdpolpk/v1
  • 1Duke Molecular Physiology Institute, Durham, NC, USA;
  • 2Department of Neurology, Duke University, Durham, NC, USA
Valerie Bekker
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DOI: dx.doi.org/10.17504/protocols.io.81wgbxdpolpk/v1
Protocol Citation: Karen Abramson, Simon Gregory 2023. U54 SCENT 10x Genomic Single-Cell Multiome. protocols.io https://dx.doi.org/10.17504/protocols.io.81wgbxdpolpk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 07, 2023
Last Modified: August 16, 2023
Protocol Integer ID: 83009
Abstract
This document outlines the 10X Genomics Single-Cell Multiome Protocol used for normal lung and colon tissues for SCENT at Duke University.
10X Gemonics Single-Cell Multiome Protocol
10X Gemonics Single-Cell Multiome Protocol
Nuclei were isolated using this 10x Genomics kit - Chromium Nuclei Isolation with RNase Inhibitor Kit, 16rxns, and this protocol

Multiome libraries were generated using these kits - Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle and this protocol

Libraries were sequenced on Illumina sequencers to a read depth of at least 50k reads/nucleus for Gene Expression with read lengths of 28x10x10x90 and at least 25k reads/nucleus for ATAC with read lengths of 50x8x24x49.