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Sequencing-based large-scale genomics approaches with small numbers of isolated maize meiocytes
High-throughput sequencing has become the large-scale approach of choice to study global gene expression and the distribution of specific chromatin marks and features. However, the limited availability of large amounts of purified cells made it very challenging to apply sequencing-based techniques i...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3933774/ https://www.ncbi.nlm.nih.gov/pubmed/24611068 http://dx.doi.org/10.3389/fpls.2014.00057 |
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author | Dukowic-Schulze, Stefanie Sundararajan, Anitha Ramaraj, Thiruvarangan Mudge, Joann Chen, Changbin |
author_facet | Dukowic-Schulze, Stefanie Sundararajan, Anitha Ramaraj, Thiruvarangan Mudge, Joann Chen, Changbin |
author_sort | Dukowic-Schulze, Stefanie |
collection | PubMed |
description | High-throughput sequencing has become the large-scale approach of choice to study global gene expression and the distribution of specific chromatin marks and features. However, the limited availability of large amounts of purified cells made it very challenging to apply sequencing-based techniques in plant meiosis research in the past. In this paper, we describe a method to isolate meiocytes from maize anthers and detailed protocols to successfully perform RNA-seq, smRNA-seq, H3K4me3-ChIP-seq, and DNA bisulfite conversion sequencing with 5000–30,000 isolated maize male meiotic cells. These methods can be adjusted for other flowering plant species as well. |
format | Online Article Text |
id | pubmed-3933774 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-39337742014-03-07 Sequencing-based large-scale genomics approaches with small numbers of isolated maize meiocytes Dukowic-Schulze, Stefanie Sundararajan, Anitha Ramaraj, Thiruvarangan Mudge, Joann Chen, Changbin Front Plant Sci Plant Science High-throughput sequencing has become the large-scale approach of choice to study global gene expression and the distribution of specific chromatin marks and features. However, the limited availability of large amounts of purified cells made it very challenging to apply sequencing-based techniques in plant meiosis research in the past. In this paper, we describe a method to isolate meiocytes from maize anthers and detailed protocols to successfully perform RNA-seq, smRNA-seq, H3K4me3-ChIP-seq, and DNA bisulfite conversion sequencing with 5000–30,000 isolated maize male meiotic cells. These methods can be adjusted for other flowering plant species as well. Frontiers Media S.A. 2014-02-25 /pmc/articles/PMC3933774/ /pubmed/24611068 http://dx.doi.org/10.3389/fpls.2014.00057 Text en Copyright © 2014 Dukowic-Schulze, Sundararajan, Ramaraj, Mudge and Chen. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Plant Science Dukowic-Schulze, Stefanie Sundararajan, Anitha Ramaraj, Thiruvarangan Mudge, Joann Chen, Changbin Sequencing-based large-scale genomics approaches with small numbers of isolated maize meiocytes |
title | Sequencing-based large-scale genomics approaches with small numbers of isolated maize meiocytes |
title_full | Sequencing-based large-scale genomics approaches with small numbers of isolated maize meiocytes |
title_fullStr | Sequencing-based large-scale genomics approaches with small numbers of isolated maize meiocytes |
title_full_unstemmed | Sequencing-based large-scale genomics approaches with small numbers of isolated maize meiocytes |
title_short | Sequencing-based large-scale genomics approaches with small numbers of isolated maize meiocytes |
title_sort | sequencing-based large-scale genomics approaches with small numbers of isolated maize meiocytes |
topic | Plant Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3933774/ https://www.ncbi.nlm.nih.gov/pubmed/24611068 http://dx.doi.org/10.3389/fpls.2014.00057 |
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