Utjecaj metode izolacije DNA na određen sastav intestinalne mikrobiote

master's thesis

• Title (english): The effect of DNA extraction method on the gut microbiota composition
• Author: Maja Ćutić
• Mentor: Jurica Žučko (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Section for Bioinformatics); Zagreb
• Defense date and country: 2017-05-09, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Nutrition

Abstract

Crijevna mikrobiota predstavlja relativno novo, zanimljivo područje istraživanja povezano s mnogim stanjima ljudskog organizma, od održavanja homeostaze preko povremenih gastrointestinalnih poremećaja, do povezanosti s bolestima. Povezanost s pretilošću omogućava bolji uvid u prevenciju, razvoj i liječenje ove bolesti čija incidencija kontinuirano raste. Kako bi se bolje razumjela ta povezanost, koriste se suvremene metode analize koje uključuju sekvencioniranje bakterijskog 16S rRNA gena iz uzorka fecesa. Cilj ovog rada bio je usporediti tri metode za izolaciju DNA iz uzorka fecesa pretile osobe kako bi se vidjele razlike u dobivenim podacima i utjecaj ovog pretkoraka na daljnje analize. Također, promatrao se utjecaj dodavanja DNA ShieldTM stabilizatora. Obradom podataka pomoću QIIME bioinformatičkog alata, dobiveni su podaci o DNA prinosu i bioraznolikosti uzoraka u ovisnosti o metodi izolacije. Za daljna istraživanja poželjno je koristiti jednu metodu kako bi se smanjila pogreška odnosno dobili sastavi mikrobiote koji su međusobno usporedivi.

Abstract (english)

Human gut microbiota is rapidly becoming one of the most interesting fields of studies regarding health and disease development. Emerging evidences show relationship between gut microbiota and obesity, one of the fastest delevoping diseases worldwide. To better understand this connection, methods such as 16S rRNA sequencing are applied. The aim of this study was to compare three available methods of DNA extraction that were used to extract DNA from faeces of a obese person. Data collected from sequencing were processed using QIIME bioinformatical tool in order to observe differences in bacterial diversity and DNA yield depending on DNA extraction method. The influence of applying DNA ShieldTM reagens was also observed. Io order to decrease biases in studies regarding analysis of the gut microbiota, it is recommended to use one specific method of DNA extraction.

• Keywords: crijevna mikrobiota; pretilost; metode izolacije DNA; 16S rRNA sekvencioniranje; QIIME program
• Keywords (english): gut microbiota; obesity; DNA isolation methods; 16S rRNA sequencing; QIIME pipeline
• Language: croatian
• URN:NBN: urn:nbn:hr:159:806794
• Study programme: Title: Nutrition; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra nutricionizma (magistar/magistra nutricionizma)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2017-11-23 10:35:30