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VOL. 6, ISSUE 4 (2021)
An optimized method for the isolation of high-quality functional RNA from polyphenol and polysaccharide-rich in vitro cultured cells of tea
Authors
Sagarika Das, Monoranjan Goswami, Rajnarayan Singh Yadav, Tanoy Bandyopadhyay
Abstract
The tea plant is rich in polyphenols which attribute to its quality and flavor. However, it also hinders the isolation of high-quality RNA from tea samples. Furthermore, tea's callus and cells in suspension cultures are rich in polysaccharides and polyphenols, which act as a bottleneck during functional RNA isolation, hampering its quality and yield immensely. In this study, a lithium chloride-based isolation procedure teamed with phenol-based extraction of RNA was optimized to overcome these hurdles. The method yielded high RNA (175.8-260.8µg/g FW) with A260/280 ranging from 1.82-2.08 and A260/230 ranging from 1.83-2.02, indicating that the isolated RNA was free of protein, polyphenol, and polysaccharide contaminants. The isolated RNA was also found to be suitable for downstream processing such as reverse transcription, PCR and qRT-PCR analysis. Therefore, this method can be used for the isolation of high-quality RNA from in vitro cultured cells of tea with high polyphenol and polysaccharide content, and also open avenues for future research in molecular biology related to callus and suspension cultures of tea.
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Pages:384-387
How to cite this article:
Sagarika Das, Monoranjan Goswami, Rajnarayan Singh Yadav, Tanoy Bandyopadhyay "An optimized method for the isolation of high-quality functional RNA from polyphenol and polysaccharide-rich <em>in vitro</em> cultured cells of tea ". International Journal of Botany Studies, Vol 6, Issue 4, 2021, Pages 384-387
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