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.
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Abbreviations
- ESTs:
-
expressed sequence tags
- dpa:
-
days postanthesis
- gfw:
-
gram fresh weight
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Wu, Y., Llewellyn, D.J. & Dennis, E.S. A quick and easy method for isolating good-quality RNA from cotton (Gossypium hirsutum L.) tissues. Plant Mol Biol Rep 20, 213–218 (2002). https://doi.org/10.1007/BF02782456
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DOI: https://doi.org/10.1007/BF02782456