Mini-prep method suitable for a plant breeding program |
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Authors: | K. M. Haymes |
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Affiliation: | (1) Centre for Plant Breeding and Reproduction Research (CPRO-DLO), Department of Vegetables and Fruit Crops, PO Box 16, 6700 AA Wageningen, The Netherlands |
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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. |
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Keywords: | DNA isolation PCR Southern blots |
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