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Validating internal controls for quantitative plant gene expression studies

validating internal controls for quantitative plant gene expression studies-82

Rice (Oryza sativa L.) is an important crop and the most advanced model for monocotyledonous species; its nuclear genome has been sequenced and molecular tools are being developed for functional analyses.However, high-throughput methods for rice research are still limited and a large-scale q RT-PCR platform for gene expression analyses has not been reported.

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Multi-parallel q RT-PCR allows the versatile and sensitive transcriptome profiling of large numbers of rice transcription factor genes.However the usefulness of these is often limited by their sensitivity and accuracy, particularly for low-abundance transcripts.In contrast, quantitative reverse transcription – polymerase chain reaction (q RT-PCR or real-time RT-PCR) allows even weakly expressed genes to be accurately quantified [2].Background Auxin/Indole-3-Acetic Acid (Aux/IAA) and Auxin Response Factor (ARF) transcription factors are key regulators of auxin responses in plants.We identified the suites of genes in the two gene families in Populus and performed comparative genomic analysis with Arabidopsis and rice.The cookies contain no personally identifiable information and have no effect once you leave the Medscape site.

Quantitative reverse transcription – polymerase chain reaction (q RT-PCR) has been demonstrated to be particularly suitable for the analysis of weakly expressed genes, such as those encoding transcription factors.

However, accurate measurement of gene expression with this method relies on the choice of a valid reference for data normalization.

Studies rarely verify that gene expression levels for reference genes are adequately consistent among the...

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Thus, whilst array-based hybridisation typically allows the detection of one transcript per cell [3, 4], q RT-PCR can detect one transcript per 1000 cells [5].