Efficient protocol for isolation of functional RNA from different grape tissue rich in polyphenols and polysaccharides for gene expression studies
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Keywords

differential display RT-PCR
grape
RNA isolation
RT-PCR
subtractive hybridization
Vitis.

How to Cite

1.
Vasanthaiah HK, Katam R, Sheikh MB. Efficient protocol for isolation of functional RNA from different grape tissue rich in polyphenols and polysaccharides for gene expression studies. Electron. J. Biotechnol. [Internet]. 2008 Jun. 15 [cited 2024 Dec. 22];11(3):0-. Available from: https://www.ejbiotechnology.info/index.php/ejbiotechnology/article/view/v11n3-5

Abstract

Extraction of RNA from plant tissue containing high levels of polyphenols and polysaccharides is tedious and difficult in grapes. Although several protocols have been published for plant RNA isolation, most have failed to yield high quality RNA in sufficient quantity from mature and diseased grape tissue. We describe a protocol for isolating intact and high quality RNA from various grape tissues as evident by high A260/A280 absorbance ratio (1.8 to 1.9) and electrophoretic profile on denaturing formaldehyde agarose gel. On an average, 205 µg RNA per g of fresh tissue were obtained using this modified protocol. RNA quality was further assessed through RT-PCR, differential display RT-PCR and subtractive hybridization, and found to be suitable for molecular studies.

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