Automated pipetting system coupled with micro- and nanoparticles as a new tool for study of nucleic acids

abstract

English

Nucleic acids still belong to the mostly studied biologically active substances. Circulating free DNAs, which were discovered at the end of forties of 20th century, demonstrate higher significance in diagnostics and study of malignant tumours. Their concentration in blood plasma significantly increases with the occurrence of malignant tumour disease. Connections with the resistance to chemotherapeutics were also demonstrated. Apoptotic cells, but also metabolically active and living cells are sources of circulating free DNA (cfDNA) in blood plasma/serum and urine. Individual studies detecting cfDNA differ according to used analytical techniques, which complicate comparison of data from individual studies. This fact obstructs real prognostic potential of this approach. Development of new methods for study of cfDNAs in patients with malignant tumour disease is of great interest. In this study, we utilized paramagnetic particles modified with nucleic acids sequences for isolation of nucleic acids. Primarily, we tested the whole approach on DNA standard and then on cfDNAs. The procedure was consequently automated by means of automated pipetting system. The isolated nucleic acids were determined electrochemically, particularly, by cyclic and square wave voltammetry. Detection limit for nucleic acids was below hundreds of ng per ml.

Pipetting, microparticles, nanoparticles, nucleic acids

MASAŘÍK, M.; HÚSKA, D.; ADAM, V.; HUBÁLEK, J.; PROVAZNÍK, I.; TRNKOVÁ, L.; KIZEK, R.

1. 1. 2010

Int. J. Mol. Med.

1107-3756

INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE

26

1

United States of America

S46

S46

1