In situ analysis of gene expression in plants.
In the post-genomic era, it is necessary to adapt methods for gene expression and functional analyses to more high-throughput levels of processing. mRNA in situ hybridization (ISH) remains a powerful tool for obtaining information regarding a gene's temporal and spatial expression pattern and can therefore be used as a starting point to define the function of a gene or a whole set of genes. We have deconstructed 'traditional' ISH techniques described for a range of organisms and developed protocols for ISH that adapt and integrate a degree of automation to standardized and shortened protocols. We have adapted this technique as a high-throughput means of gene expression analysis on wax-embedded plant tissues and also on whole-mount tissues. We have used wax-embedded wheat grains and Arabidopsis floral meristems and whole-mount Arabidopsis roots as test systems and show that it is capable of highly parallel processing.
Department of Molecular, Cell and Developmental Biology, Yale University, New Haven, CT 06520, USA.
Sinéad
S
Drea
Paul
P
Derbyshire
Rachil
R
Koumproglou
Liam
L
Dolan
John H
JH
Doonan
Peter
P
Shaw
D017360:Arabidopsis / Q000235:genetics; D035264:Flowers / Q000235:genetics; D015870:Gene Expression; D017403:In Situ Hybridization / Q000295:instrumentation / Q000379:methods*; D018517:Plant Roots / Q000235:genetics; D010944:Plants / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D018749:RNA, Plant / Q000235:genetics; D014908:Triticum / Q000235:genetics
D012333:RNA, Messenger; D018749:RNA, Plant