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.
Keywords: meiocytes, meiosis, Illumina sequencing, RNA-seq, ChIP-seq, DNA methylation, small RNA, maize
Citation: Dukowic-Schulze S, Sundararajan A, Ramaraj T, Mudge J and Chen C (2014) Sequencing-based large-scale genomics approaches with small numbers of isolated maize meiocytes. Front. Plant Sci. 5:57. doi: 10.3389/fpls.2014.00057
Received: 26 December 2013; Accepted: 05 February 2014;
Published online: 25 February 2014.
Edited by:Chung-Ju Rachel Wang, Academia Sinica, Taiwan
Reviewed by:Daphné Autran, Institut de Recherche pour le Développement, France
Copyright © 2014 Dukowic-Schulze, Sundararajan, Ramaraj, Mudge and Chen. 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.
*Correspondence: Changbin Chen, Department of Horticultural Science, University of Minnesota, 432 Alderman Hall, St. Paul, MN 55108, USA e-mail: firstname.lastname@example.org