INFLUENCE OF DIFFERENT DNA EXTRACTION METHODS ON DNA QUALITY

Deoksiribonukleinska kiselina, DNA je genetski, nasljedni materijal prisutan u stanicama organizama svih živih bića, od najjednostavnijih do najsloženijih. Obično je vrlo teško izdvojiti i pročistiti kvalitetnu DNA iz žitarica zbog pojave polisaharida, proteina i DNA, kao inhibitora polimeraze kao što su tanini, alkaloidi, te polifenoli u ekstraktima. Ciljevi ovoga istraživanja bili su utvrditi kvalitetu i koncentraciju izolirane DNA iz biljke pšenice koristeći tri različite metode izolacije. Kvaliteta izoliranih DNA izmjerena je na spektrofotometru, tako što su izmjerene apsorbance na 260 i 280 nm. Omjeri absorbanci (A260/A280) su se kretali u rasponu od 1,92 (CTAB metoda), do 1,73 (BIOBASIC metoda). Najveća koncentracija DNA dobivena je primjenom SDS metode i to 2800 ng/μl, zatim 2280 ng/μl primjenom CTAB metode i najmanja koncentracija je dobivena BIOBASIC metodom (482,5 ng/μl). Sve tri metode su se pokazale učinkovitima za izolaciju DNA iz lista biljke pšenice te se mogu koristiti ovisno o svrsi daljnjeg istraživanja.Deoxyribonucleic acid, DNA is the genetic hereditary material is present in cells of all living beings, from the simplest to the most complex. Usually it is very difficult to extract and purify high quality DNA from cereal because of presence of polysaccharides and proteins, as well as polymerase inhibitors such as tannins, alkaloids and polyphenols. The objective of this study was to determine the quality and quantity of the extracted isolated DNA from wheat seedlings using three different extraction methods, CTAB, SDS and a BIOBASIC commercial kit. The purity of the extracted DNA was measured by spectrophotometer using the absorbance at 260 and 280 nm. Absorbance ratio (A260/A280) ranged from 1.92 for CTAB method, to 1.73 for BIOBASIC method. The highest concentration of DNA was obtained using the SDS method, which yielded 2800 ng/μl, then 2280 ng/μl using CTAB method, and the lowest concentration was obtained by BIOBASIC method (482.5 ng/μl). All DNA extraction methods proved effective for DNA extraction from wheat seedlings and can be used depending on the purpose of further research