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1

New multiplex PCR assays for estimating genetic diversity in rainbow trout (Oncorhynchus mykiss) by polymorphism of microsatellite DNA

Kaczmarczyk D., Kaczor A.

Environmental Biotechnology

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2013

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Vol. 9, No. 1

19--24

EN

Multiplex PCR is a useful technique for estimating genetic diversity. This paper presents 3 new sets of primer pairs for effectively amplifying 10 microsatellite DNA loci from rainbow trout (Oncorhynchus mykiss). Unlike other sets of primer pairs that have been developed for amplifying rainbow trout microsatellite loci, ours do not require the hot-start PCR technique. In the paper, we describe the steps taken to choose the loci for each multiplex assay and to verify the genotyping results. We provide the compositions of the PCR mixture and the characteristics of the PCR thermal profile recommended for amplification.

2

Polymorphism of microsatellite loci - a tool in studying biodiversity of paddlefish aquaculture broodstock

Kaczmarczyk D., Kohlmann K., Kersten P., Luczynski M.

Environmental Biotechnology

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2007

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Vol. 3, No. 2

44-48

EN

American paddlefish (Polyodon spathula) is a new species in Polish aquaculture, its broodstocks are few and small, and it is possible that all mature fish originated from only a few spawners. Studies on polymorphism of highly variable microsatellite DNA allow revealing genetic characteristics of individual spawners as well as estimation of genetic variation within and divergence between broodstocks. This paper describes optimised protocols for isolation of DNA from fin tissues, amplification of nine microsatellite loci using PCR technique, and for fish genotyping using automatic capillary DNA sequencer. Our technique was tested towards the fin samples taken from all paddlefish reared in Poland and approaching their sexual maturity; the study included also samples taken from 47 fish of the Ukrainian breeding center (Gorny Tykich).

3

Genetic identification of black caviar based on microsatellite DNA analysis

Fopp-Bayat D.

Environmental Biotechnology

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2007

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Vol. 3, No. 2

57-60

EN

Sturgeons (Acipenseridae) are producers of black caviar, which is sold world-wide. Black caviar differs widely in quality, availability, price and taste and for these reasons it is often the subject of commercial fraud. Identification of sturgeon species is frequently based on molecular methods such as analysis of nuclear DNA and mitochondrial DNA. In this study two fragments of microsatellite DNA: Afu-39 and Afu-68 were analyzed. In caviar samples, four alleles were observed in locus Afu-39, and six alleles in locus Afu-68. This paper describes the use of microsatellite DNA markers for species identification of black caviar.

4

Microsatellite DNA polymorphism in sturgeon species and their hybrids reared in Polish aquaculture farms

Fopp-Bayat D., Luczynski M.

Environmental Biotechnology

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2006

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Vol. 2, No. 1

11-19

EN

Highly variable microsatellite DNA loci show high levels of variation per locus and provide molecular markers for species and for populations of small effective size. In this study we applied microsatellite markers to identify specimens belonging to several sturgeon species and their interspecific hybrids. Nine microsatellite loci (Afu-19, Afu-22, Afu-34, Afu-39, Afu-54, Afu-57, Afu-58, Afu-68, Afu-69) were analysed for five species (beluga, Siberian sturgeon, Russian sturgeon, sterlet and paddlefish) and for their three interspecific hybrids: Siberian sturgeon x Russian sturgeon, beluga x sterlet (called bester) and for beluga x bester. Certain alleles at five loci: Afu-22, Afu-39, Afu-54, Afu-57 and Afu-68 were diagnostic for the examined sturgeon species and their interspecific hybrids.