U54 SCENT 10x Genomic Single-Cell Multiome
Karen Abramson, Simon Gregory
Published: 2023-08-17 DOI: 10.17504/protocols.io.81wgbxdpolpk/v1
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Abstract
This document outlines the 10X Genomics Single-Cell Multiome Protocol used for normal lung and colon tissues for SCENT at Duke University.
Steps
10X Gemonics Single-Cell Multiome Protocol
1.
Nuclei were isolated using this 10x Genomics kit - Chromium
Nuclei Isolation with RNase Inhibitor Kit, 16rxns, and thisprotocoll
2.
Multiome libraries were generated using these kits -
Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle and thisprotocoll
3.
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.
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