SOLID - PHASE DNA EXTRACTION USING VARIOUS TYPES OF MAGNETIC MICROPARTICLES

abstract

English

Isolation of DNA is an important step in the molecular diagnostics of microorganisms. A high quality of isolated DNA is necessary for DNA amplification by the polymerase chain reaction. The conventional DNA isolation using phenol-chloroform extraction and DNA precipitation in ethanol is time-consuming and requires the use of toxic phenol. Magnetic separation techniques using magnetic solid particles are one of modern methods to speed up the nucleic acids isolation. The aim of of this work was to use 3 different types of magnetic microparticles, prepared by Horák, for solid - phase DNA extraction. The amounts of DNA in separation mixtures were measured by using UV-VIS spectrophotometry. The first experimental conditions were tested on chicken erythrocytes DNA. Phosphate buffer (pH 7, 7.6 and 8) was used for adsorption of magnetic particles. It was shown that approximately one half of DNA was adsorbed to the particles. The elution conditions of DNA were also optimized. Secondly bacterial DNA was tested. This DNA eluted from the particles was in PCR – ready quality.

DNA isolation, magnetic particles, PCR

KONEČNÁ, J.; ŠPANOVÁ, A.; HORÁK, D.; RITTICH, B.

2. 9. 2015

Brno University of Technology

Brno

978-80-214-5228-2

Chemistry & Life 2015 - Book of Abstracts

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