A new and simple method for rapid extraction and isolation of high-quality RNA from grape (Vitis vinifera) berries

BACKGROUND: A fast and easy method to isolate RNA from grape (Vitis vinifera) berries is proposed. RESULTS: The procedure uses high concentrations of cetyltrimethylammonium bromide (CTAB) and includes boric acid with the purpose of increasing the RNA extraction. Eppendorf tubes can be used since the amount of sample and volumes of reagents needed are small. The method is based on genomic DNA digestion with DNase I, which avoids the disadvantages of using LiCl (e.g. possible amplification of rare transcripts and reduction or even suppression of the activity of RNA-dependent DNA polymerase). The use of DNase reduces the duration of the isolation method to 8 h. The RNA yield (32.9-49.2 mu g g(-1)) and purity indices were satisfactory in all cases. CONCLUSION: Although there is a referenced grape berry method (RNeasy Midi Kit + polyethylene glycol) that is faster than the one proposed, it is 3.7 times more expensive and gives a lower yield. The ability to successfully amplify from the RNA extract was verified by measuring the beta-tubulin and lipoxygenase gene expression. (c) 2007 Society of Chemical Industry.