Optimization of polymerase chain reaction conditions for studies of paddlefish (Polyodon spathula) microsatellite DNA

Kurta, Khrystyna; Malysheva, O.; Spyrydonov, V. G.

doi:10.15407/animbiol19.02.056

Cited by 3 publications

(1 citation statement)

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“…Multiplex PCR mix with a total volume of 20.0 μl contained 50.0 mM of Tris-HCl (pH 8.3), 1.5 mM of MgCl 2 , 0.2 mM of each dNTP, 5 pM of forward and reverse primers and 1.5 U of Taq-DNA polymerase (Thermo Scientific™, Lithuania). PCR amplification (PCR) was performed on the Veriti 96 Well termocycler (Applied Biosystems, USA) by standardized parameters: (a) initial denaturation -5 min, 95 °С; (b) 30 cycles, including denaturation -15 s, 95° C; primers hybridization -25 s, 56 °С; elongation -5 s, 72 °С and (c) final elongation -5 min, 72 °С [25].…”

Section: Pcr Amplificationmentioning

confidence: 99%

Genetic variation and phylogenetic relationships among domesticated and wild paddlefish (Polyodon spathula) populations

Kurta¹,

Malysheva²,

Skrypkina

2020

Biopolym. Cell

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To determine the genetic variation and phylogenetic relationships among domesticated paddlefish populations from Ukraine and Poland with wild populations from the United States using microsatellite DNA markers. Methods. In total, 105 paddlefish individuals were analyzed using PCR technique, capillary electrophoresis, and statistical methods. Results. The evaluated mean number of alleles per locus (Na) varied from 5.5 to 6.6 in the Polish and Ukrainian paddlefish stocks, respectively, whereas twofold lower genetic polymorphism was demonstrated if compared to that found in wild population from the USA (11.1 alleles per locus). The mean observed (Ho) heterozygosity estimated in Ukrainian populations (0.709) was relatively close to those in the Polish (0.809) and wild (0.817) populations. Close genetic relationships were observed among the studied Ukrainian populations. The measured genetic similarity ranged from 0.908 (among populations from Kherson and Chernihiv regions) to 0.981 (among populations from Chernihiv and Vinnytsia regions). The Nei's genetic distances ranged from 0.019 (populations from Chernihiv and Vinnytsia regions) to 0.096 (populations from Kherson and Chernihiv regions). The constructed dendrogram demonstrated the genetic differences among hatchery-reared and natural paddlefish populations, which were separated into two single clusters according to their geographical origin. Conclusion. It has been revealed that Polish paddlefish population was more genetically distant from the studied Ukrainian populations of this species, which confirms the formation of Ukrainian and Polish populations from different parental individuals. The results obtained allow us to consider the investigated fish groups as a potential source of the exchange and enrichment of genetic resources of the paddlefish populations.

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Section: Pcr Amplificationmentioning

confidence: 99%