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However, accurate measurement of gene expression with this method relies on the choice of a valid reference for data normalization.
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 .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 .Recent improvements in q RT-PCR methodology have eliminated many of the initial problems that were associated with quantitative gene expression studies, such as those arising from alternative splicing events .
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Various high throughput techniques that allow the accurate quantification of expression levels (transcript abundance) of hundreds or thousands of genes are currently available .