Svetlana KOTOVA, Iosif TYSBOVSKY, Alena SHYLA, Nikolay KUZUB

Highly effective universal method for DNA isolation from different forensic biological traces based on the combination of adsorption chromatography on calcium-tartrate gel and gel-filtration chromatography with Toyopearl HW-65F microcolumns.

Geniş ölçekli biyolojik kalıntı- lardan DNA izolasyonu ve saf- laştırılması ile ilgili evrensel birmetod geliştirilmiştir. Bu metotard arda uygulanan iki meto- dun (Kalsiyum tartarat jelindeadsorpsiyon kromatografisi veToyopearl HW-65F mikrokolon- lu jel-filtrasyon kromatografisi)kombinasyonuna dayanmaktadır.Hematoporfirinler, kloroporfirin- ler, melaninler, humik asit, biyo- polimerler ve bunların degradas- yonuyla oluşan primer-benzeriürünler gibi kontaminantlardanayrıştırılmış yüksek saflıkta vekonsantrasyonda DNA, bu me- todla elde edilebilir.

FARKLI ADLİ BİYOLOJİK KALINTILARDAN DNA İZOLASYONU İÇİN KALSİYUM TARTARAT JELİNDE ADSORPSİYON KROMATOGRAFİSİ VE TOYOPEARL HW-65F MİKROKOLONLU JEL-FİLTRASYON KROMATOGRAFİSİ KOMBİNASYONUNA DAYALI YÜKSEK ETKİLİ EVRENSEL BİR YÖNTEM

AMAÇ: A universal method for DNA isolation and purification from the wide spectrum of biological traces has been developed. The method is based on the combination of two consequently performed methods - adsorption chromatography on calcium-tartrate gel and gel-filtration chromatography with Toyopearl HW-65F microcolumns. Highly pure concentrated DNA free of contaminants such as hematoporphyrins, chloroporphyrins, melanins, humic acids, biopolymers and products of their degradation including primer-like products can be obtained with our metho

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1. Graham DE. The isolation of high molecular weight DNA from whole organisms or large tissue masses. Anal Biochem 1978;85(2):609 13.
2. Akhrem A, Drozhdenyuk AP. Calcium tartrate gel. Anal Biochem 1989;179(1):86-9.
3. Toyopearl Instruction Manual. Tosoh Bioscience GMBH Germany 15 p Available at: http://www.separations.eu.tosohbioscience. com/NR/rdonlyres/DD2316A4-7228-420B- 908A-35E724C3369F/0/M09P18A_TIM.pdf. Cited: September, 01, 2013.
4. Maniatis T, Fritsch EF and Sambrook J. Molecular Cloning: A Laboratory Manual. New York: Cold Spring Harbor Laboratory Press, 1982.