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bioRxiv · 10.1101/2020.07.06.189506

A simple method for isolating high-quality RNA from kenaf containing high polysaccharide and polyphenol contents

Liao, X.·Li, H.·Khan, A.·Zhao, Y.·Hou, W.·Tang, X.·Akhtar, K.·Zhou, R.
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Abstract

The isolation of high-quality RNA from kenaf is crucial for genetic and molecular biology studies. However, high levels of polysaccharide and polyphenol compounds in kenaf tissues could irreversibly bind to and coprecipitate with RNA, which complicates RNA extraction. In the present study, we proposed a simplified, time-saving and low-cost extraction method for isolating high quantities of high-quality RNA from several different kenaf tissues. RNA quality was measured for yield and purity, and the proposed protocol yielded high quantities of RNA (10.1-12.9 μg/g·FW). Spectrophotometric analysis showed that A260/280 ratios of RNA samples were in the range of 2.11 to 2.13, and A260/230 ratios were in the range of 2.04-2.24, indicating that the RNA samples were free of polyphenols, polysaccharides, and protein contaminants after isolation. The method of RNA extraction presented here was superior to the conventional CTAB method in terms of RNA isolation efficiency and was more sample-adaptable and cost-effective than commercial kits. Furthermore, to confirm downstream amenability, the high-quality RNA obtained from this method was successfully used for RT-PCR, real-time RT-PCR and Northern blot analysis. We provide an efficient and low-cost method for extracting high quantities of high-quality RNA from plants that are rich in polyphenols and polysaccharides, and this method was also validated for the isolation of high-quality RNA from other plants.View Full Text

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Liao, X., Li, H., Khan, A., Zhao, Y., Hou, W., Tang, X., Akhtar, K., Zhou, R.. 2020-07-07. A simple method for isolating high-quality RNA from kenaf containing high polysaccharide and polyphenol contents. https://doi.org/10.1101/2020.07.06.189506

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