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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...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MyJove Corporation
2017
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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 |
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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 |
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