article id 1674,
category
Research note
Highlights:
We combined eleven SSR markers in one multiplex PCR to make faster and cost effective amplification of the common walnut DNA from Croatia; Genetic variation of common walnut from Croatia was moderate at analyzed SSR loci; The resultant multiplex PCR could be used for genotyping of common walnut populations.
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Multiplex polymerase chain reaction (PCR) allows amplification of two or more pair of primers in parallel for amplification of multiple target sequences in a single reaction tube. In this study, we combined existing simple sequence repeat (SSR) markers (nuclear microsatellites) in the novel combination of multiplex PCR to study the population genetics of common walnut from Croatia. From twenty one tested SSR markers, eleven produced satisfactory results in one multiplex PCR. Population genetic results achieved from 15 samples of Croatian common walnut showed moderate genetic variability (average value: He 0.473; Ho 0.568). Our multiplex PCR allowed cost effective work concerning chemicals, plastic ware, device, and working time producing optimal results. The optimized multiplex PCR represented the best combination of eleven SSR primers for genotyping common walnut in a single PCR reaction.
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Ćelepirović,
Division of Genetics, Forest Tree Breeding and Seed Science, Croatian Forest Research Institute, Jastrebarsko, Croatia
E-mail:
celepirovic.nevenka@gmail.com
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Karija Vlahović,
DNA Laboratory, Department of Forensic Medicine&Criminology, School of Medicine, University of Zagreb, Zagreb, Croatia
E-mail:
monika.karija.vlahovic@mef.hr
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Dounavi,
Department of Forest Protection, Forest Research Insitute of Baden-Württemberg, Wonnhaldesttr. 4, 79100 Freiburg, Germany
E-mail:
aikaterini.dounavi@forst.bwl.de
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Ivanković,
Division of Genetics, Forest Tree Breeding and Seed Science, Croatian Forest Research Institute, Jastrebarsko, Croatia
E-mail:
mladeni@sumins.hr