TY - JOUR
T1 - Sequencing-based large-scale genomics approaches with small numbers of isolated maize meiocytes
AU - Dukowic-Schulze, Stefanie
AU - Sundararajan, Anitha
AU - Ramaraj, Thiruvarangan
AU - Mudge, Joann
AU - Chen, Changbin
PY - 2014/2/25
Y1 - 2014/2/25
N2 - 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.
AB - 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.
KW - ChIP-seq
KW - DNA methylation
KW - Illumina sequencing
KW - Maize
KW - Meiocytes
KW - Meiosis
KW - RNA-seq
KW - Small RNA
UR - http://www.scopus.com/inward/record.url?scp=84901025257&partnerID=8YFLogxK
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U2 - 10.3389/fpls.2014.00057
DO - 10.3389/fpls.2014.00057
M3 - Review article
C2 - 24611068
AN - SCOPUS:84901025257
SN - 1664-462X
VL - 5
JO - Frontiers in Plant Science
JF - Frontiers in Plant Science
IS - FEB
M1 - 57
ER -