diplomsko delo
Vahida Puškar Šupuk (Author), Franc Janžekovič (Mentor), Nataša Pipenbaher (Co-mentor)

Abstract

V filogenetskih študijah obstajajo številne metode izolacije DNK iz živalskih celic. Namen diplomskega dela je ugotoviti ustrezno metodo izolacije DNK, ki bi nam služila za nadaljnje filogenetske analize. Primerjali smo klasično fenol - kloroform metodo izolacije DNK s komercialnim kompletom QIAamp DNA Mini Kit-om (QIAGEN). Kot model smo vzeli ribe iz rodu mren, Barbus Cuvier. Vzorci mren so bili različne starosti, zato smo jih poimenovali Skupina 1 (izlov 2011.) in Skupina 2 (izlov 2004.). DNK smo izolirali iz dela analne plavuti mren. Z elektroforezo na agaroznem gelu smo ugotavljali kakovost izolirane DNK pri obeh metodah ter med skupinama. S pomočjo Qubit fluorimetra pa smo izmerili koncentracije izoliranih vzorcev DNK. Zaradi nejasnosti, katera metoda izolacije je uspešnejša, smo nadaljevali analize s PCR metodo, s katero smo pomnožili regijo citokrom b na mtDNK. Sklepali smo, da je za izolacijo genomske DNK bolj primerna metoda s QIAamp DNK Mini Kit-om, vendar pa to ne velja za analize s PCR metodo, metodo pomnoževanja regije citokroma b na mtDNK, kjer je uspešnejša klasična metoda s fenol - kloroformom. Kakovost izolirane DNK je boljša s kompletom QIAamp DNA Mini Kitom (QIAGEN), prav tako je čas izolacije krajši s kompletom. Iz česar lahko zaključimo, da sta obe metodi primerni za izolacijo DNK, vendar je postopek s kompletom QIAamp DNA Mini Kitom (QIAGEN) enostavnejši in hitrejši.

Keywords

diplomska dela;izolacija DNK;ribe;mrene;elektroforeza;PCR metoda;

Data

Language: Slovenian
Year of publishing:
Typology: 2.11 - Undergraduate Thesis
Organization: UM FNM - Faculty of Natural Sciences and Mathematics
Publisher: [V. Puškar Šupuk]
UDC: 577.2(043.2)
COBISS: 20290568 Link will open in a new window
Views: 1715
Downloads: 139
Average score: 0 (0 votes)
Metadata: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Other data

Secondary language: English
Secondary title: Comparison of DNA isolation methods om barbels (Barbus Cuvier, 1817)
Secondary abstract: Phylogenetic studies may be performed on a wide range of DNA isolation methods. The main purpose of this graduation thesis was to determine the best DNA isolation method for possible phylogenetic analysis. The well - established phenol-chloroform method was compared to the QIAamp DNA Mini Kit (QIAGEN). Barbels, subgenus Barbus Cuvier, were the fish chosen for this comparison. The samples were divided into Group 1 (harvest in 2011) and Group 2 (harvest in 2004) to separate them according to their age. The DNA was isolated from a small portion of the anal fin. The quality of the isolated DNA was always checked with the electrophoresis on horizontal agarose gel. The concentration of the isolated DNA was then measured with Qubit fluorometer. In order to gather more data in favour of one method we have proceded with the amplification of the cytochrome b, one of the mtDNA genes, by using the PCR method. We have concluded that the QIAamp DNA Mini Kit is more suitable for the isolation of the genomic DNA, but the phenol-chloroform method proves to be better when it comes to amplification of cytochrome b. The advantages of the QIAamp Mini Kit are faster isolation and high quality of the isolated DNA. We have thus proven that both methods are suitable for DNA isolation, but the work is faster and simpler with QIAamp DNA Mini Kit.
Secondary keywords: electrophoresis;PCR;cytochrome b;barbels.;
URN: URN:SI:UM:
Type (COBISS): Undergraduate thesis
Thesis comment: Univ. v Mariboru, Fak. za naravoslovje in matematiko, Oddelek za biologijo
Pages: IX, 63 f.
ID: 8728527