SEMI-QUANTITATIVE RT-PCR ANALYSIS FOR DEHYDRIN GENE EXPRESSION LEVELS IN WHEAT UNDER DROUGHT STRESS
Abstract
Semi quantitative RT-PCR protocol was optimized in our laboratory to extract mRNA from drought stressed and control plants of two wheat cultivars and to measure the expression levels of several target mRNAs from each sample. In this investigation dehydrin gene expression was assessed. The detailed procedure for dehydrin expression levels under drought stress and control condition was described. The protocol was used as an internal control to normalize for sample to sample variations in total RNA amounts and for reaction efficiency. As for all quantitative techniques, great care must be taken in all optimization steps to ensure near quantitative (semi-quantitative) analyses. Also northern blotting for dehydrin was performed to confirm the results of semi quantitative RT-PCR in assessing the expression of gene(s) conferring dehydrin under drought stress and control condition. Generally, the results of the two techniques indicated that dehydrin gene expression increased under drought stress as compared with control plants particularly in cultivar G160.References
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