Cargando…

Integrating readout of somatic mutations in individual cells with single-cell transcriptional profiling

In many biological applications, the readout of somatic mutations in individual cells is essential. For example, it can be used to mark individual cancer cells or identify progenies of a stem cell. Here, we present a protocol to perform single-cell RNA-seq and single-cell amplicon-seq using 10X Chro...

Descripción completa

Detalles Bibliográficos
Autores principales: Liu, Shichen, Nguyen, Maximilian, Hormoz, Sahand
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8313752/
https://www.ncbi.nlm.nih.gov/pubmed/34337442
http://dx.doi.org/10.1016/j.xpro.2021.100673
_version_ 1783729410824732672
author Liu, Shichen
Nguyen, Maximilian
Hormoz, Sahand
author_facet Liu, Shichen
Nguyen, Maximilian
Hormoz, Sahand
author_sort Liu, Shichen
collection PubMed
description In many biological applications, the readout of somatic mutations in individual cells is essential. For example, it can be used to mark individual cancer cells or identify progenies of a stem cell. Here, we present a protocol to perform single-cell RNA-seq and single-cell amplicon-seq using 10X Chromium technology. Our protocol demonstrates how to (1) isolate CD34+ progenitor cells from human bone marrow aspirate, (2) prepare single-cell amplicon libraries, and (3) analyze the libraries to assign somatic mutations to individual cells. For complete details on the use and execution of this protocol, please refer to Van Egeren et al. (2021).
format Online
Article
Text
id pubmed-8313752
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-83137522021-07-31 Integrating readout of somatic mutations in individual cells with single-cell transcriptional profiling Liu, Shichen Nguyen, Maximilian Hormoz, Sahand STAR Protoc Protocol In many biological applications, the readout of somatic mutations in individual cells is essential. For example, it can be used to mark individual cancer cells or identify progenies of a stem cell. Here, we present a protocol to perform single-cell RNA-seq and single-cell amplicon-seq using 10X Chromium technology. Our protocol demonstrates how to (1) isolate CD34+ progenitor cells from human bone marrow aspirate, (2) prepare single-cell amplicon libraries, and (3) analyze the libraries to assign somatic mutations to individual cells. For complete details on the use and execution of this protocol, please refer to Van Egeren et al. (2021). Elsevier 2021-07-17 /pmc/articles/PMC8313752/ /pubmed/34337442 http://dx.doi.org/10.1016/j.xpro.2021.100673 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Liu, Shichen
Nguyen, Maximilian
Hormoz, Sahand
Integrating readout of somatic mutations in individual cells with single-cell transcriptional profiling
title Integrating readout of somatic mutations in individual cells with single-cell transcriptional profiling
title_full Integrating readout of somatic mutations in individual cells with single-cell transcriptional profiling
title_fullStr Integrating readout of somatic mutations in individual cells with single-cell transcriptional profiling
title_full_unstemmed Integrating readout of somatic mutations in individual cells with single-cell transcriptional profiling
title_short Integrating readout of somatic mutations in individual cells with single-cell transcriptional profiling
title_sort integrating readout of somatic mutations in individual cells with single-cell transcriptional profiling
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8313752/
https://www.ncbi.nlm.nih.gov/pubmed/34337442
http://dx.doi.org/10.1016/j.xpro.2021.100673
work_keys_str_mv AT liushichen integratingreadoutofsomaticmutationsinindividualcellswithsinglecelltranscriptionalprofiling
AT nguyenmaximilian integratingreadoutofsomaticmutationsinindividualcellswithsinglecelltranscriptionalprofiling
AT hormozsahand integratingreadoutofsomaticmutationsinindividualcellswithsinglecelltranscriptionalprofiling