Izolacija DNA iz bioloških tragova

master's thesis

• Title (english): DNA isolation from biological stains
• Author: Mia Knobloch
• Mentor: Milovan Kubat (mentor)
• Committee member: Vedrana Petrovečki (predsjednik povjerenstva)
• Committee member: Davor Mayer (član povjerenstva)
• Committee member: Milovan Kubat (član povjerenstva)
• Granter: University of Zagreb; School of Medicine; (Department of Forensic Medicine); Zagreb
• Defense date and country: 2015-07-15, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Clinical Medical Sciences; Forensic Medicine

Abstract

1953. godine Watson i Crick otkrili su strukturu DNA, a ubrzo potom su drugi znanstvenici otkrili da je DNA jedinstvena za svako živo biće. Zahvaljujući tome DNA se može na primjer koristiti za razlikovanje uzoraka s mjesta zločina. Uzorci mogu biti različiti, ali najčešće korišteni su krv, slina, sjemena tekućina, kosti, dlake. Oni moraju biti pažljivo prikupljeni da ne bi došlo do kontaminacije te transportirani u laboratorij kako bi dobiveni rezultati bili valjani dokaz na sudu. Četiri najčešće korištene metode izolacije u DNA laboratoriju Medicinskog fakulteta Sveučilišta u Zagrebu su organska izolacija, InstaGeneTM Matrix (Biomax), QIAamp DNA Mini kit (QIAgen) i FUJI poluautomatizirana jedinica za izolaciju DNA. Izolirana DNA se potom pohranjuje na -20°C ili -80°C. Idući korak je kvantifikacija DNA, postupak kojim se određuje kvaliteta i kvantiteta DNA iz uzorka. Potom slijedi PCR, metoda in vitro amplifikacije fragmenata DNA. Rezultat toga je mješavina amplikona koji se moraju razdvojiti kapilarnom elektroforezom ili gel elektroforezom. Na kraju postupka imamo veliki broj istih DNA fragmenata koji se mogu koristiti u istrazi.

Abstract (english)

In 1953 Watson and Crick discovered the structure of DNA molecule and soon after that scientists discovered it is unique for every living creature. Because of that it can be for example used to distinguish samples on crime scenes. Samples can be various but most commonly used are blood, saliva, semen, bones, hair. Samples must be collected carefully without contamination and transported to a laboratory so the results could be valid at the court. Four most common isolation methods in DNA laboratory at School of Medicine, University of Zagreb are organic isolation, InstaGeneTM Matrix (Biomax), QIAamp DNA Mini kit (QIAgen) i FUJI semi-automated unit for DNA extraction. Isolated DNA is then stored at -20°C or -80°C. The next step is the DNA quantification, the procedure to determine quality and quantity of the DNA from the sample, after which it is possible to do the PCR, the method of in vitro amplification of DNA fragments. The result is a mix of amplicons which needs to be separated by capillary electrophoresis or gel electrophoresis. In the end, we have a big number of the same DNA fragments which can be used in an investigation.

• Keywords: DNA izolacija; biološki tragovi; PCR; elektroforeza
• Keywords (english): DNA isolation; biological stains; PCR; electrophoresis
• Language: croatian
• URN:NBN: urn:nbn:hr:105:025270
• Study programme: Title: Medicine; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: doktor/doktorica medicine (doktor/doktorica medicine)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2016-08-12 10:20:29