magistrsko delo
Špela Buneto (Author), Tjaša Cerar (Reviewer), Jerneja Ambrožič (Mentor), Tadej Pajič (Co-mentor)

Abstract

Izolacija ustrezno čiste in kakovostne deoksiribonukleinske kisline (DNA) je predpogoj za nadaljnje analize, ki so pomembne za diagnosticiranje bolezni, napovedovanje zdravljenja, spremljanje učinka zdravljenja ter izboljšanje kakovosti življenja posameznika. Na Kliničnem inštitutu za genomsko medicino (KIGM) v Univerzitetnem kliničnem centru v Ljubljani, zagotavljajo možnosti diagnostike in genetskega svetovanja bolnikom in družinam z genetsko pogojenimi boleznimi. S strokovnim in raziskovalnim delom omogočajo prenos najsodobnejših spoznanj v vsakodnevno obravnavo genetsko pogojenih bolezni na področjih, kot so genetska epidemiologija bolezni, ki je pomembna za javno zdravstvo, moške in ženske neplodnosti in nevrogenetike, vključujoč tudi raziskovanje povezanosti mikrobioma z nevrodegenerativnimi boleznimi. Na KIGM vrsto let izvajajo osamitev DNA iz različnih bioloških vzorcev s komercialno dostopnimi reagenčnimi kompleti. Zadnja leta pa želijo povečali pretočnost izolacije DNA in razširili nabor vzorcev, iz katerih bi lahko pridobili zadostne količine kakovostne molekule DNA. Zaradi tega smo se odločili, da v sklopu magistrske naloge preverimo učinkovitost aparata ChemagicTM360, ki omogoča avtomatiziran način izolacije zelo čistih in visokokakovostnih molekul DNA iz različnih človeških vzorcev, kot tudi bakterijske DNA iz brisa ustne sluznice. Z raziskavo smo želeli preveriti, ali je izolacija z avtomatizirano metodo primerljiva z že vpeljanimi metodami izolacije. Z analizami merjenja količin, čistosti in preverjanja učinkovitosti izolacije DNA smo ugotovili, da je avtomatizirana metoda primerljiva z izbranimi, v laboratoriju že vpeljanimi, metodami izolacije in je ustrezna za uporabo v rutinski genetski diagnostiki.

Keywords

molekularna genetika;izolacija nukleinskih kislin;kri;tkivo FFPE;plodovo tkivo;cfDNA;bakterijska DNA;količina DNA;čistost DNA;učinkovitost izolacije;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL BF - Biotechnical Faculty
Publisher: [Š. Buneto]
UDC: 577.21:575.113:616-056.7
COBISS: 110764547 Link will open in a new window
Views: 115
Downloads: 24
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Other data

Secondary language: English
Secondary title: Efficiency of DNA isolation from various human tissues and of bacterial DNA from oral mucosa swabs by an automated method based on magnetic beads
Secondary abstract: Isolation of deoxyribonucleic acid (DNA) with a high level of purity and quality is a prerequisite for further analyses, which are of great importance for diagnosing human diseases, predicting treatments, monitoring the impacts of treatments, and improving the quality of life of individuals. At the Clinical Institute of Medical Genetics, a department of the University Medical Centre, they work with molecular genetic tests for the management of genetic diseases and oncological disorders, and among other things, they research the relationship between the microbiome and neurodegenerative diseases. They have established the isolation of nucleic acid from various biological samples using commercially available reagent tests for several years. In recent years they have been endeavoring to increase the flow rate of the samples, broadening the specter of samples from which they can collect DNA and improve the quality of the isolated nucleic acids. Therefore, we decided that validation of the ChemagicTM360 instrument should be a part of my master's thesis. This instrument has an automated system for DNA purification from various samples of human tissue as well as bacterial DNA from oral cavity swabs. As part of the ongoing research, we wanted to verify that the automated method was equivalent to well-established laboratory methods for isolation of nucleic acid. By analysing the quantity, purity, and verifying the efficiency of nucleic acid isolation, we concluded that the validated method can be substituted for reference isolation methods and is suitable for use in routine diagnostics.
Secondary keywords: molecular genetics;nucleic acid isolation;blood;FFPE tissue;fetal tissue;cfDNA;bacterial DNA;amount of DNA;quantity of DNA;efficiency of isolation;
Type (COBISS): Master's thesis/paper
Study programme: 0
Thesis comment: Univ. v Ljubljani, Biotehniška fak., Oddelek za mikrobiologijo
Pages: XIII f., 74 str., [21] str. pril.
ID: 15595149