20.500.12556/DKUM-18955 Primerjava različnih metod izolacije DNA pri bezgu (Sambucus spp.) Comparison of different DNA extraction methods from elderberry leaves (Sambucus spp.) Cilj študije je bil ugotoviti, katere izolacije DNA so kvalitativno, časovno in stroškovno najbolj smotrne za uporabo. V raziskave smo vključili štiri različne metode izolacije DNA, in sicer klasično CTAB metodo ter tri metode izolacije s kompleti: Wizard Genomic DNA Purification Kit (Promega), Total DNA Purification Kit (Invitrogen) in avtomatsko metodo izolacije z napravo QIAcube (Qiagen) in kompletom DNeasy plant Mini Kit (Qiagen). Vzorce bezga (Sambucus spp.) smo v letih 2010 in 2011 izolirali v laboratoriju za genetiko Fakultete za kmetijstvo in biosistemske vede Univerze v Mariboru. Izhodiščni material bezga je bil iz narave, iz rastlinjaka in iz tkivnih kultur. Izolirano DNA smo pogledali na agaroznem gelu in fluorometrično izmerili koncentracijo DNA. Po statistični obdelavi podatkov smo ugotovili, da metoda izolacije statistično značilno vpliva na kakovost in donos izolirane DNA, hkrati pa sta kakovost in donos povezana s stroški izolacije. Pri dražjih metodah izolacije je izolirana DNA bolj kakovostna, donos izolirane DNA pa nižji. Časovno je izolacija DNA z že pripravljeni kompleti precej krajša, stroškovno pa dražja v primerjavi z uveljavljeno in pogosto uporabljeno metodo CTAB. The main goal of the study was to determine which DNA extraction methods are in terms of quality, time and cost the most appropriate. Four different DNA extraction methods were included in the research, namely classical CTAB extraction method and three extraction methods with purification kits (Wizard Genomic DNA Purification Kit (Promega), Total DNA Purification Kit (Invitrogen), and automated extraction method with QIAcube (Qiagen) machine and with DNeasy plant Mini Kit (Qiagen)). DNA extraction from elderberrys was carried out in 2010 and 2011 at the Faculty of Agriculture and Life Sciences at University of Maribor in the laboratory for genetics. The original material of elderberry leaves was taken from nature, greenhouse and from tissue culture. The extracted DNA was checked in agarose gel and DNA concentration was fluorometrically measured. Statistical tests showed that the method of isolation significantly influenced the quality and yield of extracted DNA. Also the quality and yield were related with extraction costs. More expensive extraction methods resulted with higher quality and with lower yield of DNA. Concerning the time used, methods with purification kits were much quicker, but from the viewpoint of costs they were more expensive in comparison to CTAB method. izolacija DNA/ bezeg/ Sambucus spp. DNA extraction/ elderberry/ Sambucus spp. true true false Slovenski jezik Angleški jezik Diplomsko delo 2011-06-13 08:00:02 2011-06-27 10:51:36 2022-04-13 14:57:27 2023-12-24 03:31:08 2011 0 Maribor 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 URN:SI:UM:DK:U8K92DYR UNI_Leopold_Eva_2011.pdf UNI_Leopold_Eva_2011.pdf 1 F9FB0A63E9B33B57C84243A27D9E89BB 2111660cec738de4125cdd7b6e77b389273c29c6640c01757aa14db5b4f0044a 437486c3-7c05-11eb-bb7a-00155d0001ca 20.500.12556/dkum/9a51185a-ae7d-4f34-bc26-f27f7f60d655 https://dk.um.si/Dokument.php?lang=slv&id=22758 Fakulteta za kmetijstvo in biosistemske vede 0 0 0