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Mini-prep method suitable for a plant breeding program
Authors:K M Haymes
Institution:(1) Centre for Plant Breeding and Reproduction Research (CPRO-DLO), Department of Vegetables and Fruit Crops, PO Box 16, 6700 AA Wageningen, The Netherlands
Abstract:Plant breeders need a nondestructive and inexpensive protocol to screen large numbers of plants early after sowing. Isolation of DNA represents one of the limiting steps in this process: normally a person is capable of isolating 25 to 50 samples per day. To speed up the isolation of DNA in marker-assisted plant breeding programs, a CTAB mini-prep was modified into an inexpensive and nondestructive procedure. With this protocol a single individual can isolate 200 to 250 samples per day of high-quality DNA that is immediately suitable for testing by PCR. The amounts of DNA isolated are sufficient for about 150 PCR amplifications. These improvements are achieved by eliminating time-consuming and noncritical steps in the standard protocol, such as extensive grinding, washing with ethanol, and treatment with RNAse A.
Keywords:DNA isolation  PCR  Southern blots
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