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Sequencing-based large-scale genomics approaches with small numbers of isolated maize meiocytes.

Frontiers in plant science (2014-03-13)
Stefanie Dukowic-Schulze, Anitha Sundararajan, Thiruvarangan Ramaraj, Joann Mudge, Changbin Chen
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.

MATERIALS
Product Number
Brand
Product Description

Roche
Protector RNase Inhibitor
Sigma-Aldrich
Anti-trimethyl-Histone H3 (Lys4) Antibody, Upstate®, from rabbit