Naslov
Significance of application of verified sets for DNA isolation in molecular diagnostics

Autori
Kalmar Kristina ; Stipanović Tea ; Glavaš-Obrovac Ljubica ; Mihaljević Ivan

Izvornik
Acta clinica Croatica. Supplement (0353-9474) 46 (2007); 105-105

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, sažetak, znanstveni

Ključne riječi
DNA isolation ; molecular diagnostic

Sažetak
Introduction: For the molecular diagnostic it is important to obtain integral DNA without potential inhibitors of polymerase chain reaction (PCR) method. Aim: To investigate the efficiency of DNA isolation and quality of DNA, obtained from whole EDTA-blood using two commercially procurable sets for DNA isolation, as well as standard phenol- chloroform method. Materials and methods: DNA was isolated using "High Pure PCR-Template Preparation Kit" (Roche, Germany, EU) - set 1 and "Biotools Bloodclean DNA Purification Kit" (Biotools, Spain, EU) - set 2, as well as phenol-chloroform DNA extraction method. Fifty EDTA-blood samples were analyzed. Results: Time required for DNA isolation using commercial sets 1 and 2 (45 minutes and 90 minutes per sample, respectively) compared to phenol-chloroform method (3 days per sample) was significantly shorter. Also, commercial sets were less toxic in comparison to standard method. Set 1 was the most reliable because obtained DNA was intact and its concentration was within precisely defined range. Application of set 2 resulted in a high content of broken, useless DNA. Besides, by using DNA samples, obtained by set 2, PCR reaction was inhibited, due to residual chemicals in 20% of cases. Conclusion: Since quality of commercially procurable sets for DNA isolation considerably differs, it is very important to examine the quality of obtained DNA, as well as hurtfulness of sets, and choose a test with the best characteristics for molecular diagnostic purpose.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Temeljne medicinske znanosti

Napomena
Abstract of the Scientific Syposium with International Participation "40 Years of Nuclear Medicine in Osijek", November 15 - 17, 2007. Osijek, Croatia.

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