Abstract | Deoksiribonukleinska kiselina (Deoxyribonucleic Acid, DNA) je središnja molekula života te osnovni nositelj nasljedne informacije, kontrolira rast i razvoj svakog živog bića. Oplemenjivački programi koriste izoliranu DNA u istraživanjima genetske različitosti, u stvaranju novih kultivara, kultivara s većim prinosom, otpornijih na bolesti i štetnike. U radu je korištena Cetiltrimetilamonijev bromid (CTAB) metoda. Ciljevi ovog istraživanja su utvrditi kvalitetu i količinu izolirane DNA iz biljke kukuruza, pšenice, suncokreta, uljane repice, lana bundeve i krastavca. Čistoća dobivene DNA testirana je spektrofotometrom, a na temelju dobivenih vrijednosti izračunata je koncentracija DNA. Spektrofotometrijskom analizom dobivene su vrijednosti absorbanci (A260nm i A280nm), a omjer je bio u rasponu od 1,7 (lan) do 1,89 (uljana repica). Omjeri absorbanci ispod 1,7 dobiveni su za dvije kulture kukuruz (1,41) i bundevu (1,36) te su uzorci označeni kao nečisti. Korištena metoda pokazala se učinkovita za izolaciju DNA iz , pšenice, suncokreta, uljane repice, lana i krastavca dok je za kukuruz i bundevu potrebno metodu dodatno prilagoditi. |
Abstract (english) | Deoxyribonucleic Acid or DNA is a basic life molecule, the most important carrier of heredity. The DNA controls growth and development of every living being. Breeding programs use this information that DNA gives in various research regarding genetic diversity, creating of new cultivars, cultivars with higher yield, good quality and resistance to pests and diseases. widely accepted Cetyl trimethylammonium bromide (CTAB) method is used in this research. Aims of this research were to determine quality and concentration of extracted DNA from maize, wheat, sunflower, olidseed rape, flax, pumpkin and cucumber. Quality of extracted DNA was estimated by spectrophotometer, and based on those measurements DNA concentration was calculated. Absorbance values (260 nm and 280 nm) ranged from 1.7 (for flax) till 1.89 (for oilseed rape). Absorbance ratio (A260/A280) lower than 1.7 was estimated in two DNA samples: maize (1.41) and pumpkin (1.36), and were classified as impure. CTAB method was very effective for five tested plant species (wheat, sunflower, olidseed rape, flax and cucumber) while further method modifications should be done for maize and pumpkin. |