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United States Department of Agriculture

Agricultural Research Service

Title: Optimization of Microsatellite Methodologies for the Analysis of Pecan Variation

Authors
item Mendoza-Herrera, A - TEXAS A&M UNIVERSITY
item Grauke, Larry
item Loopstra, C - TEXAS A&M UNIVERSITY

Submitted to: Plant and Animal Genome VX Conference Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: November 15, 1999
Publication Date: N/A

Technical Abstract: Microsatellites or Simple Sequences Repeats (SSR) were demonstrated to be a powerful tool for studying characterization, conservation, and improvement of important agricultural crops. The great value of these markers arises because they are hypervariable, co-dominant, abundant, reproducible, and easy to detect by PCR method. At the moment, not much information is found don Pecan [Carya illinoinensis (Wangenh.) K. Koch], a very important nut tree crop in North America. For that, this work was conducted to optimize and establish the most convenient methodology with high levels of confidence and reproducibility but with the lowest cost for the development of the Pecan microsatellites results. DNA samples of 15 cultivars of different geographic origin were amplified by PCR using 5 pairs of selected SSR primers. The amplification was carried out using P-32 labeled primers and non-labeled primers. The PCR products were electrophoresed on 6% denaturing acrylamide gels and 3% and 4% agarose gels. Both visualization systems were compared in the resolution of the banding pattern. The agarose gel systems were selected and different agarose gel compositions were tested in order to obtain the best separation and resolution of the SSR bands. Metaphor agarose, a combination of Metaphor with normal agarose and Synergel (polysaccharide product) with normal agarose were compared and the results with each one will be discussed. To reduce the number of PCR reactions/sample, number of gels, time and money, PCR with two different pair of SSR primers was conducted and very good results were found with some primer combinations. Samples from a wider geographical range are currently being evaluated with additional SSR primers and these analyses and results will also be discussed.

Last Modified: 11/28/2014
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