470 PB 339 DNA AMPLIFICATION USING TAQ POLYMERASE AND STOFFEL FRAGMENT IN POLYPLOID SWEETPOTATO

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  • 1 Dept. of Horticulture, LSU Agricultural Center, Baton Rouge, LA 70803
  • | 2 Dept. of Agronomy, LSU Agricultural Center, Baton Rouge, LA 70803

RAPD and the single dose polymorphic band (SDPB) are powerful tools for genome map construction of higher polyploids, such as hexaploid sweetpotato. Duplication in the genome of higher polyploids results in fewer polymorphisms per primer screened than one would expect in diploids. The Stoffel fragment (Sf) is suggested as an alternative to the most commonly used Taq DNA polymerase to maximize the number of polymorphisms. Genomic DNA from two sweetpotato varieties, `Excel' and `Beauregard', and F1 progeny was isolated using a modified CTAB procedure. The DNA was assayed with twelve primers from Operon Technologies groups A and F. Each enzyme was tested with and without a ramp temperature treatment between the annealing and the extension temperatures. Results are based on three separate amplifications and electrophoretic runs. Band reproducibility was better using Sf than Taq; unfortunately, resolution was lower making bands difficult to score. 8.4% more scorable bands and 20.3% more storable polymorphisms were obtained with Taq. The ramp treatment did not alter results using Sf, but did improve the reproducibility of Taq and ease scoring. The number of bands and their location were the same.

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