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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...

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Autores principales: Dukowic-Schulze, Stefanie, Sundararajan, Anitha, Ramaraj, Thiruvarangan, Mudge, Joann, Chen, Changbin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2014
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.
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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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