High-throughput, high-sensitivity analysis of gene expression in Arabidopsis.
High-throughput gene expression analysis of genes expressed during salt stress was performed using a novel multiplexed quantitative nuclease protection assay that involves customized DNA microarrays printed within the individual wells of 96-well plates. The levels of expression of the transcripts from 16 different genes were quantified within crude homogenates prepared from Arabidopsis (Arabidopsis thaliana) plants also grown in a 96-well plate format. Examples are provided of the high degree of reproducibility of quantitative dose-response data and of the sensitivity of detection of changes in gene expression within limiting amounts of tissue. The lack of requirement for RNA purification renders the assay particularly suited for high-throughput gene expression analysis and for the discovery of novel chemical compounds that specifically modulate the expression of endogenous target genes.
Kris
Richard Martin
RM
NuvoGen Research LLC, Tucson, AZ 85728, USA. rkris@nuvogenresearch.com
Felder
Stephen
S
Deyholos
Michael
M
Lambert
Georgina M
GM
Hinton
James
J
Botros
Ihab
I
Martel
Ralph
R
Seligmann
Bruce
B
Galbraith
David W
DW
D000040:Abscisic Acid / Q000494:pharmacology; D000222:Adaptation, Physiological; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression; D020869:Gene Expression Profiling / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D012965:Sodium Chloride / Q000494:pharmacology
D012965:Sodium Chloride; D000040:Abscisic Acid