DNA fragment analysis by agarose gel electrofhoresis method

Abstract

Agarozna gel elektroforeza je migracija nabijenih čestica na gelu, uslijed djelovanja električnog polja. Uobičajena je laboratorijska tehnika za odvajanje i analizu fragmenata DNA i ostalih biomolekula na temelju njihove veličine. Genomska DNA je ukupan genetički materijal jednog organizma i nalazi se u jezgrama stanica. Cilj ovog rada bio je opisati i primijeniti metodu agarozne gel elektroforeze te utvrditi veličine fragmenata DNA molekule. Pripremljene su četiri vrste gelova koncentracija: 0,8%, 1%, 2% i 3%. Prema dobivenim rezultatima utvrđeno je da su fragmenti genomske DNA veći od PCR fragmenata i sukladno s tim je zaključeno da manji fragmenti putuju brže i dalje od velikih fragmenata, što je bilo i očekivano. Zaključno, za analizu većih fragmenata genomske DNA optimalno je koristiti gel koncentracije 1% i manje, dok se za PCR fragmente najčešće koristi gel kocentracije od 2 do 3%, optimalno 2,5%.Agarose gel electrophoresis is the migration of charged particles on the gel, due to the influence of applied electric field. It is a common laboratory technique for separating and analyzing DNA fragments and other biomolecules based on their size. Genomic DNA is a total genetic material of an organism and it's found in the cell nuclei. The aim of this research was to describe and apply the agarose gel electrophoresis method and to determine the sizes of DNA molecule fragments. Four gel concentrations were prepared: 0.8%, 1%, 2% and 3%. According to the obtained results, it was determined that genomic DNA fragments are larger than PCR fragments and accordingly it was concluded that smaller fragments travel faster than the large fragments, which was expected. In conclusion, for the analysis of larger fragments of genomic DNA, it is optimal to use a gel concentration of 1% or less, while for PCR fragments, gel concentration of 2 to 3% is usually used, optimally 2,5%