A versatile nuclei extraction protocol for single cell multiome ATAC and gene expression in non model species v1

Citation

Ruiz Daniels R A versatile nuclei extraction protocol for single cell multiome ATAC and gene expression in non model species v1. Refers to: 10.17504/protocols.io.bp2l69wnrlqe/v1. PLoS ONE. https://doi.org/10.17504/protocols.io.bp2l69wnrlqe/v1

Abstract
Here we present a modified version of : dx.doi.org/10.17504/protocols.io.261genwm7g47/v2 that was used to successfully extract nuclei from an array of different tissue types for single cell sequencing and modify it with the purpose of extracting nuclei for single cell multiome ATAC and gene expression on the 10x chromium. The modifications for this protocol include: different concentration of RNase inhibitors, different quantities for nuclear isolation buffer, removal of unnecessary steps as well as QC specific for multiome analysis. If you are looking to use this protocol for bulk ATAC-seq use of protease inhibitor cocktail PIC is recommended instead of RNase inhibitor on the snRNA-seq version of this protocol (dx.doi.org/10.17504/protocols.io.261genwm7g47/v2) please get in touch with the authors if you are unsure on how to do this.

People

Dr Rose Ruiz Daniels

Lecturer in Aquaculture Genomics, Institute of Aquaculture