Author/Editor		Zabavnik-Piano, J; Uršič, M
Title		A simple method for isolation of DNA from various samples
Translated title		Preprosta metoda za izolacijo DNK iz različnih vzorcev
Type		članek
Source		Zb Vet Fak Univ Ljublj
Vol. and No.		Letnik 33, št. 2
Publication year		1996
Volume		str. 185-98
Language		eng
Abstract		Various methods have been specifically designed to isolate deoxyribonucleic acid (DNA) from different sources. In order to use only one method to isolate DNA from different origins we have modified the method originally described to purify microbial nuclieic acids from samples containing only small quantities of cells. We are successfully using this method to isolate genomic DNA of high purity from skin and variously treated blood samples. The procedure uses guanidinium thiocyanate (GuSCN) and its potential to lyse cells and inactive endogenous nucleases. In the presence of high concentration of this chaotropic agent nuclecic acids bind to diatoms, glass or silica particles. Although, Boom et.al. (3) do not recommend using silica particles to isolate nucleic acids from cell-rich specimens in our experience the obtained DNA was always of good quality to perform enzymatic reactions like polymerase chain reaction (PCR) amplification and restriction digestions. The method is fast, simple and reliable, does not need special equipment and is specially suited for the diagnostic laboratories that only occasionally isolate genomic material from varous tissue samples.
Summary		Poznanih je mnogo specifičnih za izolacijo deoksiribonukleinskih kislin (DNK) iz različnih vzorcev. Da bi z eno samo metodo lahko izolirali DNK iz različnih vzorcev, smo v našem laboratoriju modificirali metodo, ki je bila prvotno opisana za prečiščevanje nukleinskih kislin mikrobov iz celično revnih vzorcev. To metodo uspešno uporabljamo za icolacijo genomske DNK iz kože različno obdelainaktivira endogene nukleaze. Poleg tega se prvi visokih koncentracijah GuSCN nukleinske kisline vežejo na delce stekla ali slike. Čeprav Boom in sod. (3) ne priporočajo uporabe silikonskih delcev za izolacijo nukleinskih kislin iz vzorcev, bogatih s celicami, pa naše izkušnje kažejo, da je s to metodo dobljena DNK primarna za nadaljnje encimsko obdelovanje: za pomnoževanje s polimerazno verižno reakcijo (PCR) in za razrez DNK z restrikcijskimi encimi. Metoda je hitra, preprosta in zanesljiva. Ker ni potrebna posebna oprema, je zelo primerna za diagnostične laboratorije, v katerih le občasno izolirajo genomski material iz različnih tkivnih vzorcev.
Descriptors		DNA SWINE POLYMERASE CHAIN REACTION GUANIDINES SILICON DIOXIDE