Please use this identifier to cite or link to this item:https://hdl.handle.net/20.500.12259/52775
Type of publication: Tezės kituose recenzuojamuose leidiniuose / Theses in other peer-reviewed publications (T1e)
Field of Science: Biologija / Biology (N010)
Author(s): Juškaitytė, Erika;Jocienė, Lina;Paulauskas, Algimantas;Kupčinskienė, Eugenija
Title: Selection of dominant markers for investigation of genetic diversity of Cucurbitaceae family species
Is part of: The vital nature sign [electronic resource] : 10th international scientific conference, May 19-20, 2016 Vilnius, Lithuania : abstract book. Kaunas : Vytautas Magnus university, 2016, [no. 10]
Extent: p. 44-44
Date: 2016
Keywords: Genetic diversity;Cupressaceae;ISSR;Inter-simple sequence repeat markers;Molecular markers
Abstract: In addition to cultivated for food and forage representatives of Cucurbitaceae family, some species belonging to Echinocystis, Bryonia, Ecballium and Sicyos are either cultivated for ornamental purposes or growing wild in Lithuania. The objective of present study was to select dominant Inter Simple Sequence Repeat (ISSR) markers for evaluation of genetic diversity of ornamental species Echinocystis lobata and Sicyos angulata. The total DNA was extracted by DNA Purification Kit (#KO512, Thermo Scientific, Lithuania). Three randomly chosen ISSR markers (UBC 810, UBC 881 and UBC 890) were selected and conditions suitable for DNA amplification were screened. Amplification reactions were carried out in volumes of 20 μl containing of 10 μl PCR Master mix (0.05 U/μl Taq DNA Polymerase; 4 mM MgCl2; 0.4 mM each dNTP), 0.4 μl 20 mg/ml BSA (0.6 μl of BSA was also tested), 0.5 μl 10 μM primer and 80 ng of DNA (40 ng of DNA was also tested). Two different polymerase chain reaction (PCR) programs were tested. The first one consisted of an initial denaturation at 94 °C for 5 min, followed by 35 cycles of denaturation at 94 °C for 30 s, annealing at 55 °C for 45 s, extension at 72 °C for 45 s and a final extension at 72 °C for 10 min. According to this program, none of the fragments of DNA were amplified. The second type PCR protocol included an initial denaturation at 94 °C for 5 min, followed by 45 cycles of denaturation at 95 °C for 45 s, annealing at 54–56 °C for 45 s, extension at 72 °C for 2 min and a final extension at 72 °C for 5 min, the following program allowed to amplify DNA fragments with all selected markers, although for separate primers annealing temperatures were different. To obtain polymorphic and reproducible fragments of DNA the most favorable annealing temperature for the marker UBC 810 was 54 °C and for the markers UBC 881, UBC 890 temperature was 56 °C. [...]
Internet: http://vns.microsep.org/wp-content/uploads/2016/06/vns2016VK.pdf
Affiliation(s): Biologijos katedra
Gamtos mokslų fakultetas
Vytauto Didžiojo universitetas
Appears in Collections:Universiteto mokslo publikacijos / University Research Publications

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