DNA amplification fingerprinting of plant genomes

G. Caetano-Anolles, P. M. Gresshoff

Research output: Contribution to journalArticle

Abstract

Multiple arbitrary amplicon profiling (MAAP) describes a general strategy for the analysis of simple and complex genomes that uses one or more arbitrary oligonucleotide primers to direct the enzymatic amplification of an anonymous DNA template. DNA amplification fingerprinting (DAF) uses very short (≥5 nt) primers to produce characteristic and highly informative DNA patterns that are adequately resolved by polyacrylamide gel electrophoresis and silver staining. DNA polymorphisms are useful as genetic markers in genetic mapping applications, either directly or as sequence-characterized amplified regions (SCAR). Complex patterns are ideally suited for general DNA fingerprinting. Multiple endonuclease digestion of template DNA or amplification products can increase significantly the detection of polymorphic DNA, Coupled with isogenic lines or bulked segregant analysis, markers linked to specific chromosomal regions can be effectively identified. While the first 8 nt from the 3' terminus of the primer usually direct the amplification reaction, the length of this arbitrary domain can be reduced by adding an extraordinarily stable mini-hairpin at its 5' terminus. These mini-hairpin oligonucleotides can be used to fingerprint subgenomic fragments such as cloned DNA or polymerase chain reaction-amplified DNA fragments. The application of MAAP to the analysis of plant genomes will be discussed.

Original languageEnglish (US)
Pages (from-to)62-70
Number of pages9
JournalMethods in Molecular and Cellular Biology
Volume5
Issue number1
StatePublished - Jan 1 1994
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Biology
  • Clinical Biochemistry
  • Cell Biology

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