Reproducibility of random amplified polymorphic DNA (RAPD) analysis among laboratories

Greg A. Penner, Aria Bush, Roger Wise, Won Kim, Les Domier, Ken Kasha, Andre Laroche, Graham Scoles, Stephen J. Molnar, George Fedak

Research output: Contribution to journalArticle

Abstract

Random amplified polymorphic DNA (RAPD) analysis appears to offer a cost- and time-effective alternative to restriction fragment-length polymorphism (RFLP) analysis. However, concerns about the ability to compare RAPD results from one laboratory to another have not been addressed effectively. DNA fragments that were amplified by five primers and shown to be reproducibly polymorphic between two oat cultivars (within the Ottawa laboratory) were tested in six other laboratories in North America. Four of the six participants amplified very few or no fragments using the Ottawa protocol. These same participants were able to generate a considerable number of amplified fragments by using their own protocols. The reproducibility of results among laboratories was affected by two factors. First, different laboratories amplified different size ranges of DNA fragments, and, consequently, small and large polymorphic fragments were not always reproduced. Second, although reproducible results were obtained with four of the primers, reproducible results were not obtained with the fifth primer, using the same reaction conditions. It is suggested that if the overall temperature profiles (especially the annealing temperature) in-side the tubes are identical among the laboratories, then RAPD fragments are likely to be reproducible.

Original languageEnglish (US)
Pages (from-to)341-345
Number of pages5
JournalGenome Research
Volume2
Issue number4
DOIs
StatePublished - Jan 1 1993

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

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    Penner, G. A., Bush, A., Wise, R., Kim, W., Domier, L., Kasha, K., Laroche, A., Scoles, G., Molnar, S. J., & Fedak, G. (1993). Reproducibility of random amplified polymorphic DNA (RAPD) analysis among laboratories. Genome Research, 2(4), 341-345. https://doi.org/10.1101/gr.2.4.341