Multiplex titration RT-PCR: rapid determination of gene expression patterns for a large number of genes.
We have developed an improved method for determination of gene expression levels with RT-PCR. The procedure is rapid and does not require extensive optimization or densitometric analysis. Since the detection of individual transcripts is PCR-based, small amounts of tissue samples are sufficient for the analysis of expression patterns in large gene families. Using this method, we were able to rapidly screen nine members of the Aux/IAA family of auxin-responsive genes and identify those genes which vary in message abundance in a tissue- and light-specific manner. While not offering the accuracy of conventional semi-quantitative or competitive RT-PCR, our method allows quick screening of large numbers of genes in a wide range of RNA samples with just a thermal cycler and standard gel analysis equipment.
Nebenfuhr
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Department of Botany and Plant Pathology, Oregon State University, Corvallis 97331-2902, USA.
Lomax
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D018076:DNA, Complementary; D003624:Darkness; D005069:Evaluation Studies as Topic; D018506:Gene Expression Regulation, Plant / Q000235:genetics*; D017343:Genes, Plant; D007210:Indoleacetic Acids; D008027:Light; D018551:Solanum lycopersicum / Q000235:genetics* / Q000254:growth & development; D010937:Plant Growth Regulators / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D018749:RNA, Plant; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D018076:DNA, Complementary; D007210:Indoleacetic Acids; D010937:Plant Growth Regulators; D012333:RNA, Messenger; D018749:RNA, Plant; C030737:indoleacetic acid