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Optimizacija in uporabnost metode za identifikacijo cianobakterij v okoljskih vzorcih na osnovi DNABurger, Evgenija ...Cvetenje cianobakterij je pogosto povezano z izločanjem toksinov, zato je pomembno, da pravočasno identificiramo morebitne strupene vrste cianobakterij. Cilja našega raziskovanja sta bila priprava ... hitrega, zdravju neškodljivega in učinkovitega protokola za izolacijo DNA iz okoljskih vzorcev oziroma cianobakterijskih celic ter postavitev smernic za identifikacijo posameznih cianobakterijskih vrst na osnovi analize DNA. Optimizirali smo protokol za hrambo okoljskih vzorcev, izolacijo DNA in pomnoževanje regij rRNA 16 S in ITS-1 na kromosomu. Večjo natančnost kot z določitvijo velikosti pomnoženih regij smo pri določanju heterogenosti cianobakterijskih vzorcev dosegli z restrikcijsko analizo PCR-produktov in določitvijo nukleotidnega zaporedja. Iz nabora dostopnih encimov smo kot najbolj uporabne restrikcijske nukleaze identificirali EcoRI, NcoI, EcoRV, KpnI in BclI. Pri določanju nukleotidnih zaporedij smo ugotovili, da so začetni oligonukleotidi, namenjeni za pomnoževanje genomskih regij pri cianobakterijah, pomnožili tudi nekatera kloroplastna zaporedja evkariontskih alg. Pripravili smo zgledne primere restrikcijske analize pomnoženih genomskih regij za identifikacijo dveh cianobakterij. Za vzorec z Blejskega jezera smo ugotovili, da je bilo nukleotidno zaporedje regije ITS-1 identično kot pri sevu Planktothrix rubescens NIVA_CYA18 in še nekaterih drugih. To kaže na minimalne razlike v sicer šibko ohranjeni genomski regiji ITS med različnimi izolati oziroma na svetovno razširjenost med seboj zelo podobnih sevov vrste P. rubescens.Source: Zbornik povzetkov (Str. 18)Type of material - conference contributionPublish date - 2018Language - slovenianCOBISS.SI-ID - 96346113
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Burger, Evgenija | |
Marzel Djuranovič, Lucija | |
Petravić, Luka | |
Gačnik, Tanja | |
Juteršek, Mojca, biokemičarka | 51987 |
Dolinar, Marko, biokemik | 06416 |
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