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MARIS: Method for Analyzing RNA following Intracellular Sorting
Transcriptional profiling is a key technique in the study of cell biology that is limited by the availability of reagents to uniquely identify specific cell types and isolate high quality RNA from them. We report a Method for Analyzing RNA following Intracellular Sorting (MARIS) that generates high...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3940959/ https://www.ncbi.nlm.nih.gov/pubmed/24594682 http://dx.doi.org/10.1371/journal.pone.0089459 |
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author | Hrvatin, Siniša Deng, Francis O'Donnell, Charles W. Gifford, David K. Melton, Douglas A. |
author_facet | Hrvatin, Siniša Deng, Francis O'Donnell, Charles W. Gifford, David K. Melton, Douglas A. |
author_sort | Hrvatin, Siniša |
collection | PubMed |
description | Transcriptional profiling is a key technique in the study of cell biology that is limited by the availability of reagents to uniquely identify specific cell types and isolate high quality RNA from them. We report a Method for Analyzing RNA following Intracellular Sorting (MARIS) that generates high quality RNA for transcriptome profiling following cellular fixation, intracellular immunofluorescent staining and FACS. MARIS can therefore be used to isolate high quality RNA from many otherwise inaccessible cell types simply based on immunofluorescent tagging of unique intracellular proteins. As proof of principle, we isolate RNA from sorted human embryonic stem cell-derived insulin-expressing cells as well as adult human β cells. MARIS is a basic molecular biology technique that could be used across several biological disciplines. |
format | Online Article Text |
id | pubmed-3940959 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-39409592014-03-06 MARIS: Method for Analyzing RNA following Intracellular Sorting Hrvatin, Siniša Deng, Francis O'Donnell, Charles W. Gifford, David K. Melton, Douglas A. PLoS One Research Article Transcriptional profiling is a key technique in the study of cell biology that is limited by the availability of reagents to uniquely identify specific cell types and isolate high quality RNA from them. We report a Method for Analyzing RNA following Intracellular Sorting (MARIS) that generates high quality RNA for transcriptome profiling following cellular fixation, intracellular immunofluorescent staining and FACS. MARIS can therefore be used to isolate high quality RNA from many otherwise inaccessible cell types simply based on immunofluorescent tagging of unique intracellular proteins. As proof of principle, we isolate RNA from sorted human embryonic stem cell-derived insulin-expressing cells as well as adult human β cells. MARIS is a basic molecular biology technique that could be used across several biological disciplines. Public Library of Science 2014-03-03 /pmc/articles/PMC3940959/ /pubmed/24594682 http://dx.doi.org/10.1371/journal.pone.0089459 Text en © 2014 Hrvatin et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Hrvatin, Siniša Deng, Francis O'Donnell, Charles W. Gifford, David K. Melton, Douglas A. MARIS: Method for Analyzing RNA following Intracellular Sorting |
title | MARIS: Method for Analyzing RNA following Intracellular Sorting |
title_full | MARIS: Method for Analyzing RNA following Intracellular Sorting |
title_fullStr | MARIS: Method for Analyzing RNA following Intracellular Sorting |
title_full_unstemmed | MARIS: Method for Analyzing RNA following Intracellular Sorting |
title_short | MARIS: Method for Analyzing RNA following Intracellular Sorting |
title_sort | maris: method for analyzing rna following intracellular sorting |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3940959/ https://www.ncbi.nlm.nih.gov/pubmed/24594682 http://dx.doi.org/10.1371/journal.pone.0089459 |
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