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United States Department of Agriculture

Agricultural Research Service

Title: Modified Protocols for Rapid Extraction of Plant Genomic Dna, Aflp Analysisand Visualization with Radioisotope Or Silver Stain in Carrots

Authors
item Briard, M - INST NATL HORT FRANCE
item Le Clerc, V - INST NATL HORT FRANCE
item Grzebelus, D - KRAKOW AGRI UNIV POLAND
item Senalik, D - DEPT HORT UW MADISON WI
item Simon, Philipp

Submitted to: Plant Molecular Biology Reporter
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: January 25, 2001
Publication Date: N/A

Interpretive Summary: Molecular genetic markers are important in plant breeding and genetic resources management. However, for evaluation of large numbers of samples, standard methods are often time-consuming and expensive. The AFLPtm technique is frequently used for the identification of molecular markers, owing to some important advantages over other techniques, i.e. high reproducibility, and relative technical simplicity. The use of radioactive isotopes to visualize AFLPtm products renders the technique relatively expensive, inaccessible to researchers not equipped for use of radioactivity and is hazardous to health. The technique we describe involves a simple, reliable extraction of carrot DNA, which has been problematic, AFLPtm analysis, and easy visualization. The modifications described are especially useful when large numbers of plant samples are to be studied, as they can speed up the analyses and minimize expense. With this information scientists doing molecular fingerprinting and mapping of plant genomes can gather data more rapidly accurately, and safely than with existing methods.

Technical Abstract: AFLPtm is currently one of the most frequently techniques used for the identification of molecular markers. We report certain modifications to the genomic DNA extraction, AFLP, and staining protocols, which speed up analyses and minimize expenses while screening large numbers of plant samples. Using the presented protocol we achieved 82 percent reduction of costs without compromising the reliability and quality of data gathered for carrot.

Last Modified: 8/30/2014
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