title: Sequencing-based large-scale genomics approaches with small numbers of isolated maize meiocytes.
abstract: 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.
authors.affiliation: Department of Horticultural Science, University of Minnesota St. Paul, MN, USA.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Dukowic-Schulze
authors.affiliation: National Center for Genome Resources Santa Fe, NM, USA.
authors.forename: Anitha
authors.initials: A
authors.lastname: Sundararajan
authors.affiliation: National Center for Genome Resources Santa Fe, NM, USA.
authors.forename: Thiruvarangan
authors.initials: T
authors.lastname: Ramaraj
authors.affiliation: National Center for Genome Resources Santa Fe, NM, USA.
authors.forename: Joann
authors.initials: J
authors.lastname: Mudge
authors.affiliation: Department of Horticultural Science, University of Minnesota St. Paul, MN, USA.
authors.forename: Changbin
authors.initials: C
authors.lastname: Chen
keywords: ChIP-seq; DNA methylation; Illumina sequencing; RNA-seq; maize; meiocytes; meiosis; small RNA