Dissociation of chick embryonic tissue for FACS and preparation of isolated cells for genome-wide downstream assays
Summary: In order to process samples by fluorescence-activated cell sorting (FACS), it is essential to obtain a single-cell suspension of dissociated cells. Numerous protocols and commercial reagents are available; however, each requires optimization for specific tissue types. Here, we describe an o...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-06-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721001210 |
| _version_ | 1819043168003293184 |
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| author | Ruth M. Williams Tatjana Sauka-Spengler |
| author_facet | Ruth M. Williams Tatjana Sauka-Spengler |
| author_sort | Ruth M. Williams |
| collection | DOAJ |
| description | Summary: In order to process samples by fluorescence-activated cell sorting (FACS), it is essential to obtain a single-cell suspension of dissociated cells. Numerous protocols and commercial reagents are available; however, each requires optimization for specific tissue types. Here, we describe an optimized protocol for dissociating dissected chick embryos across a broad span of developmental stages. We also provide protocols for processing targeted cell populations isolated using FACS for ATAC-seq, RNA-seq, and chromatin immunoprecipitation.For complete details on the use and execution of this protocol, please refer to Ling and Sauka-Spengler (2019) and Williams et al. (2019). |
| first_indexed | 2024-12-21T09:52:29Z |
| format | Article |
| id | doaj.art-6fa10209529a49feb076efe826d5ae27 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-21T09:52:29Z |
| publishDate | 2021-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-6fa10209529a49feb076efe826d5ae272022-12-21T19:08:08ZengElsevierSTAR Protocols2666-16672021-06-0122100414Dissociation of chick embryonic tissue for FACS and preparation of isolated cells for genome-wide downstream assaysRuth M. Williams0Tatjana Sauka-Spengler1University of Oxford, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, Oxford OX3 9DS, UK; Corresponding authorUniversity of Oxford, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, Oxford OX3 9DS, UK; Corresponding authorSummary: In order to process samples by fluorescence-activated cell sorting (FACS), it is essential to obtain a single-cell suspension of dissociated cells. Numerous protocols and commercial reagents are available; however, each requires optimization for specific tissue types. Here, we describe an optimized protocol for dissociating dissected chick embryos across a broad span of developmental stages. We also provide protocols for processing targeted cell populations isolated using FACS for ATAC-seq, RNA-seq, and chromatin immunoprecipitation.For complete details on the use and execution of this protocol, please refer to Ling and Sauka-Spengler (2019) and Williams et al. (2019).http://www.sciencedirect.com/science/article/pii/S2666166721001210GenomicsModel OrganismsMolecular BiologySequencingSingle Cell |
| spellingShingle | Ruth M. Williams Tatjana Sauka-Spengler Dissociation of chick embryonic tissue for FACS and preparation of isolated cells for genome-wide downstream assays STAR Protocols Genomics Model Organisms Molecular Biology Sequencing Single Cell |
| title | Dissociation of chick embryonic tissue for FACS and preparation of isolated cells for genome-wide downstream assays |
| title_full | Dissociation of chick embryonic tissue for FACS and preparation of isolated cells for genome-wide downstream assays |
| title_fullStr | Dissociation of chick embryonic tissue for FACS and preparation of isolated cells for genome-wide downstream assays |
| title_full_unstemmed | Dissociation of chick embryonic tissue for FACS and preparation of isolated cells for genome-wide downstream assays |
| title_short | Dissociation of chick embryonic tissue for FACS and preparation of isolated cells for genome-wide downstream assays |
| title_sort | dissociation of chick embryonic tissue for facs and preparation of isolated cells for genome wide downstream assays |
| topic | Genomics Model Organisms Molecular Biology Sequencing Single Cell |
| url | http://www.sciencedirect.com/science/article/pii/S2666166721001210 |
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