Author/Authors :
Yılmaz, Mete Ege Üniversitesi - Su Ürünleri Fakültesi - Yetiştiricilik Bölümü, Turkey , Yılmaz, Mete University of Florida - School of Forest Resources and Conservation - Program in Fisheries and Aquatic Sciences, USA , Phlips, Edward J. University of Florida - School of Forest Resources and Conservation - Program in Fisheries and Aquatic Sciences, USA
Title Of Article :
Design and Testing of Internal Amplification Controls (IAC) and Development of a Restriction Fragment Length Polymorphism (RFLP) Assay for Cylindrospermopsin Genes
Abstract :
Environmental DNA may contain impurities that inhibit downstream molecular applications. Use of an internal amplification control (IAC) is one way to detect inhibition of the polymerase chain reaction. We describe a strategy to design these amplification controls for the most commonly tested cylindrospermopsin genes cyrB and cyrC. We demonstrate PCR inhibition from DNA of samples collected from lakes Dora, Griffin, Crescent, Buffalo Bluff and George in Florida, USA by use of these internal controls. Further cleaning of DNA from impurities resulted in detection of cyrB and cyrC genes in samples from lakes Crescent, Buffalo Bluff and George. Comparison of restriction digests of cyrB fragments from lake samples, Aphanizomenon ovalisporum and Cylindrospermopsis raciborskii suggested the former cyanobacterium as the likely source of cylindrospermopsin in these samples.
NaturalLanguageKeyword :
Aphanizomenon ovalisporum , Cylindrospermopsis , cylindrospermopsin , internal amplification control , restriction fragment length polymorphism.
JournalTitle :
Ege Journal Of Fisheries and Aquatic Sciences
