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A Non-destructive Seed Sampling Method for PCR-based Analyses in Marker Assisted Selection and Transgene Screening

Permanent URL:
http://handle.nal.usda.gov/10113/14279
File:
Download [PDF File]
Abstract:
Extraction of quality DNA from peanut (Arachis hypogaea L.) generally requires extensive manipulation in order to remove numerous phenolic compounds and polysaccharides. To reduce the amount of problematic compounds present, routine purification of peanut DNA is normally performed on young leaf tissue which requires time and space for seed germination and growth of the peanut plant for at least two to four weeks. Here, we describe a simple, non-destructive method for extracting genomic DNA from a mature dry peanut seed that is of suitable quality for PCR-based molecular analysis. The method we developed requires only 0.02 g of peanut cotyledon tissue taken directly from the distal end of the mature dry seed and provides 30-46 μg of DNA suitable for use in restriction digests, PCR, SCAR and SSR analyses. This method is the first of its kind developed for DNA extraction from peanut seed. Since this method is non-destructive, seed can be subsequently germinated to produce healthy mature plants, making this technique a useful tool for the application of marker assisted selection in screening segregating populations of putative transgenic seed and in the advancement of breeding populations.
Author(s):
Chenault, K.D. , Gallo, M. , Seib, J.C. , James, V.A.
Subject(s):
Arachis hypogaea , peanuts , polymerase chain reaction , transgenes , marker-assisted selection , seeds , cotyledons , DNA , extraction , nondestructive methods , microsatellite repeats
Format:
p. 38-43.
Note:
Includes references
Source:
Peanut science 2007 Jan-June, v. 34, issue 1
Language:
English
Year:
2007
Collection:
Journal Articles, USDA Authors, Peer-Reviewed
Rights:
Works produced by employees of the U.S. Government as part of their official duties are not copyrighted within the U.S. The content of this document is not copyrighted.