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...
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| Format: | Article |
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MyJoVE Corporation
2018
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| Online Access: | http://hdl.handle.net/1721.1/115000 https://orcid.org/0000-0003-0649-7428 https://orcid.org/0000-0002-0989-8115 |
| _version_ | 1826194296813912064 |
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| author | Knouse, Kristin Ann Hendricks, Austin A Wu, Jie |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Knouse, Kristin Ann Hendricks, Austin A Wu, Jie |
| author_sort | Knouse, Kristin Ann |
| collection | MIT |
| 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. Her e 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. |
| first_indexed | 2024-09-23T09:53:52Z |
| format | Article |
| id | mit-1721.1/115000 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T09:53:52Z |
| publishDate | 2018 |
| publisher | MyJoVE Corporation |
| record_format | dspace |
| spelling | mit-1721.1/1150002022-09-30T17:34:20Z Detection of Copy Number Alterations Using Single Cell Sequencing Knouse, Kristin Ann Hendricks, Austin A Wu, Jie Massachusetts Institute of Technology. Department of Biology Koch Institute for Integrative Cancer Research at MIT Knouse, Kristin Ann Hendricks, Austin A Wu, Jie 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. Her e 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. National Institute of General Medical Sciences (U.S.) (Award T32GM007753) National Cancer Institute (U.S.) (Grant P30-CA14051) National Institutes of Health (U.S.) (Award GM056800) 2018-04-27T17:42:32Z 2018-04-27T17:42:32Z 2017-02 2018-02-23T17:22:49Z Article http://purl.org/eprint/type/JournalArticle 1940-087X http://hdl.handle.net/1721.1/115000 Knouse, Kristin A. et al. “Detection of Copy Number Alterations Using Single Cell Sequencing.” Journal of Visualized Experiments 120 (February 2017): e55143 © 2017 Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License https://orcid.org/0000-0003-0649-7428 https://orcid.org/0000-0002-0989-8115 http://dx.doi.org/10.3791/55143 Journal of Visualized Experiments Attribution-NonCommercial-NoDerivs 3.0 Unported (CC BY-NC-ND 3.0) https://creativecommons.org/licenses/by-nc-nd/3.0/ application/pdf MyJoVE Corporation Journal of Visualized Experiments |
| spellingShingle | Knouse, Kristin Ann Hendricks, Austin A Wu, Jie 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 |
| url | http://hdl.handle.net/1721.1/115000 https://orcid.org/0000-0003-0649-7428 https://orcid.org/0000-0002-0989-8115 |
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