Cargando…

Detection of Copy Number Alterations Using Single Cell Sequencing

Detection of genomic changes at single cell resolution is important for characterizing genetic heterogeneity and evolution in normal tissues, cancers, and microbial populations. Traditional methods for assessing genetic heterogeneity have been limited by low resolution, low sensitivity, and/or low s...

Descripción completa

Detalles Bibliográficos
Autores principales: Knouse, Kristin A., Wu, Jie, Hendricks, Austin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5409325/
https://www.ncbi.nlm.nih.gov/pubmed/28287554
http://dx.doi.org/10.3791/55143
_version_ 1783232457208758272
author Knouse, Kristin A.
Wu, Jie
Hendricks, Austin
author_facet Knouse, Kristin A.
Wu, Jie
Hendricks, Austin
author_sort Knouse, Kristin A.
collection PubMed
description Detection of genomic changes at single cell resolution is important for characterizing genetic heterogeneity and evolution in normal tissues, cancers, and microbial populations. Traditional methods for assessing genetic heterogeneity have been limited by low resolution, low sensitivity, and/or low specificity. Single cell sequencing has emerged as a powerful tool for detecting genetic heterogeneity with high resolution, high sensitivity and, when appropriately analyzed, high specificity. Here we provide a protocol for the isolation, whole genome amplification, sequencing, and analysis of single cells. Our approach allows for the reliable identification of megabase-scale copy number variants in single cells. However, aspects of this protocol can also be applied to investigate other types of genetic alterations in single cells.
format Online
Article
Text
id pubmed-5409325
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher MyJove Corporation
record_format MEDLINE/PubMed
spelling pubmed-54093252017-05-12 Detection of Copy Number Alterations Using Single Cell Sequencing Knouse, Kristin A. Wu, Jie Hendricks, Austin J Vis Exp Genetics Detection of genomic changes at single cell resolution is important for characterizing genetic heterogeneity and evolution in normal tissues, cancers, and microbial populations. Traditional methods for assessing genetic heterogeneity have been limited by low resolution, low sensitivity, and/or low specificity. Single cell sequencing has emerged as a powerful tool for detecting genetic heterogeneity with high resolution, high sensitivity and, when appropriately analyzed, high specificity. Here we provide a protocol for the isolation, whole genome amplification, sequencing, and analysis of single cells. Our approach allows for the reliable identification of megabase-scale copy number variants in single cells. However, aspects of this protocol can also be applied to investigate other types of genetic alterations in single cells. MyJove Corporation 2017-02-17 /pmc/articles/PMC5409325/ /pubmed/28287554 http://dx.doi.org/10.3791/55143 Text en Copyright © 2017, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Genetics
Knouse, Kristin A.
Wu, Jie
Hendricks, Austin
Detection of Copy Number Alterations Using Single Cell Sequencing
title Detection of Copy Number Alterations Using Single Cell Sequencing
title_full Detection of Copy Number Alterations Using Single Cell Sequencing
title_fullStr Detection of Copy Number Alterations Using Single Cell Sequencing
title_full_unstemmed Detection of Copy Number Alterations Using Single Cell Sequencing
title_short Detection of Copy Number Alterations Using Single Cell Sequencing
title_sort detection of copy number alterations using single cell sequencing
topic Genetics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5409325/
https://www.ncbi.nlm.nih.gov/pubmed/28287554
http://dx.doi.org/10.3791/55143
work_keys_str_mv AT knousekristina detectionofcopynumberalterationsusingsinglecellsequencing
AT wujie detectionofcopynumberalterationsusingsinglecellsequencing
AT hendricksaustin detectionofcopynumberalterationsusingsinglecellsequencing