|Kubelik Anne R|
Submitted to: Current Genetics
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/20/1995
Publication Date: N/A
Citation: Interpretive Summary: Random amplified polymorphic DNAs (RAPDs) are a recently developed method for genetic analysis of organisms. The advantages of this method include its ease and its speed; however, a major limitation is in the amount of information it provides. We have developed a modification of this technique that alleviates this limitation for the study of fungi and expands its usefulness. Because this modification is applicable to a broad range of fungi, it will be of benefit both to scientists doing basic research on fungi, as well as to those studying fungal diseases of plant and animals.
Technical Abstract: Major limitations of the RAPD technique include the proportion of primers that do not work and the low number of amplification products generated per primer. This paper describes the use of primers with G+C contents of 80-100% in RAPDs performed on fungal DNA. In analysis on Puccinia graminis f. sp. tritici DNAs, these primers generated twice the number of amplification products per primer and more than twice the number of polymorphisms between isolates as compared to 60-70% G+C primers. These RAPD markers segregate as dominant markers and do not appear to be clustered in the genome. Further analysis demonstrated that high-GC primers worked equally well in a broad range of other fungi.