Isolation and analysis of genomic DNA from single seeds
1990
Kamalay, J.C. | Tejwani, R. | Rufener, G.K. II.
Our objective was to adapt a plant DNA minipreparation procedure for the rapid isolation of total nucleic acid from single legume seeds. Single-seed extractions from cultivars of soybean (Glycine max L. Merr.) and frenchbean (Phaseolus vulgaris L.) were characterized for total nucleic acid yield, DNA content, and restriction fragment length polymorphisms (RFLPs). This procedure can be employed in the analysis of a large number of individual seeds by genomic blotting and molecular hybridization. While probing seed lots with the same complementary DNA (cDNA) fragment, we found that ostensibly pure soybean seed lines contained significant levels of identical RFLPs while frenchbean seed lines from plants with dramatic morphological differences had no detectable RFLPs for the same sequence. When isolated genomic DNAs were used to detect restriction fragment length polymorphisms (RFLPs), it was possible to make limited distinctions among cultivars of both soybean and frenchbean seed lots. This technique met the criteria of simplicity, rapidity, and low cost, and yielded DNA fragments of sufficient purity and length for RFLP analysis.
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