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A quick and easy method for isolating good-quality RNA from cotton (Gossypium hirsutum L.) tissues

A quick and easy method for isolating good-quality RNA from cotton (Gossypium hirsutum L.) tissues Conventional RNA extraction methods have been shown to produce poor quality RNA with low yields from cotton tissues. We present a protocol for quick isolation of cotton RNA with high yield and good quality. By combining a borate extraction buffer developed by Wan and Wilkins (1994) with Qiagen RNeasy spin columns, plus proteinase K treatment during extraction, we produced RNA from 0 dpa (day postanthesis) cotton ovules at an average yield of 1000 μg/gfw (gram fresh weight). The poly A+ RNA derived from these RNA preparations was capable of efficient in vitro translation and reverse transcription. This method could also be applied to other cotton tissues, including leaves, 7 dpa fibres, 7 dpa ovules, petals, hypocotyls, and roots, to produce RNA at yields ranging from 300–1000 μg/gfw with little or no degradation. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Molecular Biology Reporter Springer Journals

A quick and easy method for isolating good-quality RNA from cotton (Gossypium hirsutum L.) tissues

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References (10)

Publisher
Springer Journals
Copyright
Copyright © 2002 by Springer
Subject
Life Sciences; Plant Sciences; Plant Breeding/Biotechnology; Proteomics; Metabolomics; Bioinformatics
ISSN
0735-9640
eISSN
1572-9818
DOI
10.1007/BF02782456
Publisher site
See Article on Publisher Site

Abstract

Conventional RNA extraction methods have been shown to produce poor quality RNA with low yields from cotton tissues. We present a protocol for quick isolation of cotton RNA with high yield and good quality. By combining a borate extraction buffer developed by Wan and Wilkins (1994) with Qiagen RNeasy spin columns, plus proteinase K treatment during extraction, we produced RNA from 0 dpa (day postanthesis) cotton ovules at an average yield of 1000 μg/gfw (gram fresh weight). The poly A+ RNA derived from these RNA preparations was capable of efficient in vitro translation and reverse transcription. This method could also be applied to other cotton tissues, including leaves, 7 dpa fibres, 7 dpa ovules, petals, hypocotyls, and roots, to produce RNA at yields ranging from 300–1000 μg/gfw with little or no degradation.

Journal

Plant Molecular Biology ReporterSpringer Journals

Published: Sep 3, 2012

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