20.500.12556/DKUM-81846 Vpliv načina izolacije DNA iz svežih in liofiliziranih listov slive (Prunus domestica L.) na uporabnost DNA za nadaljnje genetske analize Effect of Different DNA Isolation Methods from Fresh and Lyophilized Plum Leaves (Prunus domestica L.) on the DNA Utility for Further Genetic Analysis V letu 2020 smo na Fakulteti za kmetijstvo in biosistemske vede v Mariboru (laboratorij za genetiko) opravili poskus, kjer smo proučevali vpliv petih različnih metod izolacije DNA (izolacija s CTAB iz svežih in liofiliziranih listov, izolacija s CTAB in dodanim PIA reagentom, izolacija z Genomic DNA Purification Kit - Monarch® ter izolacija z GenEluteTM Plant Genomic DNA Miniprep Kit - Sigma®) iz listov slive (Prunus domestica L.) na kakovost izolirane DNA. Rezultati so pokazali, da ima način izolacije statistično značilen vpliv na čistost in koncentracijo DNA. Najčistejša DNA je bila izolirana s kompletom Sigma, pri katerem je povprečna vrednost spektrofotometrije odstopala od optimalnega razmerja A260 / A280 le za 0,17. Med izolacijami DNA po metodi CTAB je bila najvišja povprečna koncentracija DNA izmerjena pri izolaciji s CTAB in dodanim PIA reagentom (486,4 ng/µL), najnižja pa pri izolaciji s CTAB iz liofiliziranih listov (256,5 ng/µL). Pri izolaciji DNA s kompletom Monarch je bila izmerjena višja povprečna koncentracija (18,09 ng/µL) kot pri kompletu Sigma (8,92 ng/µL). In the experiment which was carried out in year 2020 at the Faculty of Agriculture and Life Sciences of Maribor (Laboratory of Genetics), the effect of five different DNA isolation methods (isolation with CTAB from fresh and lyophilized leaves, isolation with CTAB and added PIA reagent, isolation with Genomic DNA Purification Kit - Monarch® and isolation with GenEluteTM Plant Genomic DNA Miniprep Kit - Sigma®) on the quality of isolated DNA from plum leaves (Prunus domestica L.) was studied. The results indicate that the isolation method has a statistically significant effect on DNA purity and concentration. The purest DNA was isolated with the Sigma kit, in which the average value of spectrophotometry deviated from the optimal A260 / A280 ratio only by 0.17. Among the isolations with the CTAB method, the highest average concentration of DNA was measured in the isolation with CTAB and added PIA reagent (486,4 ng/µL) and the lowest with CTAB from lyophilized leaves (256,5 ng/µL). In the isolation with the Monarch kit, a higher average concentration (18,09 ng/µL) was measured than with the Sigma kit (8,92 ng/µL). izolacija DNA CTAB Prunus domestica čistost DNA koncentracija DNA DNA isolation CTAB Prunus domestica DNA purity DNA concentration true true false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2022-06-08 10:23:48 2022-07-20 13:14:50 2022-08-10 00:55:20 0000-00-00 00:00:00 2022 0 Maribor 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 112913667 MAG_Senveter_Nika_2022.pdf MAG_Senveter_Nika_2022.pdf 1 B5B7945D0098BAF5776E4ADCF59AE408 edcd3496335ffa4f46425a9d60ee61c9308a520fe25631247940e4ceb9d8c113 4ed5fe17-f5ee-11ec-88fd-00155d000105 20.500.12556/dkum/d1061c77-ddb6-4d7f-b68f-128ce4882215 https://dk.um.si/Dokument.php?lang=slv&id=158954 Fakulteta za kmetijstvo in biosistemske vede 0 0 0