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
Abstract
Geniş ölçekli biyolojik kalıntılardan DNA izolasyonu ve saflaştırılması ile ilgili evrensel bir metod geliştirilmiştir. Bu metot ard arda uygulanan iki metodun Kalsiyum tartarat jelinde adsorpsiyon kromatografisi ve “Toyopearl HW-65F” mikrokolonlu jel-filtrasyon kromatografisi kombinasyonuna dayanmaktadır. Hematoporfirinler, kloroporfirinler, melaninler, humik asit, biyopolimerler ve bunların degradasyonuyla oluşan primer-benzeri ürünler gibi kontaminantlardan ayrıştırılmış yüksek saflıkta ve konsantrasyonda DNA, bu metodla elde edilebilir.
Keywords
adli biyolojik kalıntılar adsorpsiyon kromatografisi kalsiyum tartarat jel jel-filtrasyon kromatografisi Toyopearl HW-65F mikrokolon DNA izolasyonu ve saflaştırılmas
References
- Graham DE. The isolation of high molecular weight DNA from whole organisms or large tissue masses. Anal Biochem 1978;85 2 :609– 13.
- Akhrem A, Drozhdenyuk AP. Calcium tartrate gel. Anal Biochem 1989;179 1 :86-9.
- 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.
- Maniatis T, Fritsch EF and Sambrook J. Molecular Cloning: A Laboratory Manual. New York: Cold Spring Harbor Laboratory Press, 1982.
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
Abstract
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 gelfiltration 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 method.
Keywords
forensic biological traces adsorption chromatography calcium-tartrate gel gelfiltration chromatography Toyopearl HW-65F microcolumns DNA isolation and purification
References
- Graham DE. The isolation of high molecular weight DNA from whole organisms or large tissue masses. Anal Biochem 1978;85 2 :609– 13.
- Akhrem A, Drozhdenyuk AP. Calcium tartrate gel. Anal Biochem 1989;179 1 :86-9.
- 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.
- Maniatis T, Fritsch EF and Sambrook J. Molecular Cloning: A Laboratory Manual. New York: Cold Spring Harbor Laboratory Press, 1982.
Details
| Primary Language | Turkish |
|---|---|
| Subjects | Forensic Biology |
| Journal Section | Research Article |
| Authors | |
| Submission Date | January 1, 2014 |
| Publication Date | April 30, 2014 |
| IZ | https://izlik.org/JA93NA75ZX |
| Published in Issue | Year 2014 Volume: 28 Issue: 1 |
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