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Preparation of total RNA from a very small wheat embryo suitable for differential display

Chen, Y. and Weining, S. and Daggard, G. (2003) Preparation of total RNA from a very small wheat embryo suitable for differential display. Annals of Applied Biology, 143 (2). pp. 261-264.

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Organisation URL: http://www.aab.org.uk/contentok.php?id=117

Abstract

A simple method was developed for extracting total RNA from a single mature wheat seed embryo, which is very small and hard. Guanidine thiocyanate and chloroform were employed and the grinding of the samples was performed in a microcentrifuge tube with a plastic pestle. A "jacket" of liquid nitrogen and simplified procedures were applied to ensure the thorough grinding of the embryo tissue and to minimise the loss of samples. These measures substantially increased the recovery of total RNA in the extraction process. Reliable differential display was successfully achieved with the total RNA after DNase treatment and reverse transcription. This method makes it viable to study gene expression and gene regulation in a single wheat seed embryo. It may also give researchers the ability to analyse mRNAs in tissues or organs which were previously too small for RNA isolation using conventional procedures.

Item Type:Article
Additional Information:Reproduced with permission from Association of Applied Biologists. Access to published version may be available via Publisher’s website.
Keywords:Total RNA; single wheat embryo; differential display; reverse transcription.
Subjects:Plant culture > Seeds. Seed technology
Deposited On:28 Jan 2004
Last Modified:15 Nov 2010 02:55

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