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 mug/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 mug/gfw with little or no degradation.