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.
Drea
Sinéad
S
Department of Molecular, Cell and Developmental Biology, Yale University, New Haven, CT 06520, USA.
Derbyshire
Paul
P
Koumproglou
Rachil
R
Dolan
Liam
L
Doonan
John H
JH
Shaw
Peter
P
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