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I. Becker, K. Becker, M. Röhrl, H. Höfler (1997)
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The maize handbook
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Meiosis-specific gene discovery in plants: RNA-Seq applied to isolated Arabidopsis male meiocytesBMC Plant Biology, 10
Stefanie Dukowic-Schulze, A. Harris, Junhua Li, A. Sundararajan, J. Mudge, E. Retzel, W. Pawlowski, Changbin Chen (2014)
Comparative transcriptomics of early meiosis in Arabidopsis and maize.Journal of genetics and genomics = Yi chuan xue bao, 41 3
Christophe Lambing, F. Franklin, C. Wang (2017)
Understanding and Manipulating Meiotic Recombination in Plants[OPEN]Plant Physiology, 173
Changbin Chen, E. Retzel (2013)
Analyzing the meiotic transcriptome using isolated meiocytes of Arabidopsis thaliana.Methods in molecular biology, 990
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Stefanie Dukowic-Schulze, A. Sundararajan, Thiruvarangan Ramaraj, J. Mudge, Changbin Chen (2014)
Sequencing-based large-scale genomics approaches with small numbers of isolated maize meiocytesFrontiers in Plant Science, 5
A. Eveland, A. Goldshmidt, M. Pautler, K. Morohashi, Christophe Liseron-Monfils, M. Lewis, Sunita Kumari, S. Hiraga, Fang Yang, Erica Unger-Wallace, Andrew Olson, S. Hake, E. Vollbrecht, E. Grotewold, D. Ware, D. Jackson (2014)
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I. Golubovskaya (1994)
A Smear Technique for the Study of Meiosis in Pollen Mother Cells of Maize
Meiosis is essential during sexual reproduction to generate haploid gametes. Genomic or epigenomic studies of meiosis in multicellular organisms using next‐generation sequencing (NGS) methods have been limited because of the difficulty of collecting thousands to millions of meiocytes. Here, we describe a simple protocol to efficiently isolate maize male meiocytes from formaldehyde‐fixed samples for NGS techniques that require chemical crosslinking to preserve complex interactions or chromatin architecture. Anthers at desired meiotic stages are selected, fixed with paraformaldehyde, and disrupted using a homogenizer. Cell walls are digested to produce a cell suspension containing small somatic cells and large individual meiocytes. The meiocyte fraction is enriched by size separation with cell strainers and further purified by flow cytometry. From 400 anthers, we can isolate 20,000 meiocytes at 98% purity in 6 to 8 hours. © 2018 by John Wiley & Sons, Inc.
Current Protocols in Plant Biology – Wiley
Published: Jun 1, 2018
Keywords: ; ; ; ; ;
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