Isolation of High Quality RNA from Oilseeds of Jatropha curcas |
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Authors: | Rajesh K Singh Aparna Misra Vidhu A Sane Pravendra Nath |
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Institution: | 1. Plant Gene Expression Lab, National Botanical Research Institute, Lucknow, 226 001, India
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Abstract: | High quality RNA with good yield is a prerequisite for carrying out several molecular biology studies. Recalcitrant tissues such as oilseeds pose several problems while isolating good quality RNA. We have standardized a fast and simple protocol for RNA isolation from the seeds of Jatropha curcas, which gives good quality RNA without compromising for the yield. By including pre wash of seed powder with acetone and removal of polysaccharides through selective precipitation, we have been regularly isolating good quality total RNA in the range of 300–450 μg g?1 depending upon tissue type. The RNA isolated by this procedure is devoid of any contaminating DNA. The RNA preparations have been subjected to cDNA synthesis and PCR, and found suitable for these studies. This method also works satisfactorily with groundnut and mustard seeds. |
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