BIOANALYTICAL APPLICATIONS OF POLYOXOMETALATES USING MOLECULAR TOOLS FOR DETECTION OF DNA AND SNPS IN PRIMER EXTENSION

Abstract Ahmed Mehdi Debela

This PhD work was started with the intention of developing platform for electrochemical detection of mutations using highly specific method called arrayed primer extension. So the work involves the preparation of labels to be linked with the four different DNA bases and the preparation of robust platform which can withstand the high temperature during enzymatic incorporation of the labelled DNA bases. The first part of this work deals of the preparation of compounds called polyoxometalates (Keggin and Dawson types) and their covalent attachment to DNA sequence and also, to DNA single bases along with other organic redox labels for mutation detection. In the second part while trying to achieve a thermally stable surface tethered DNA probes, a new method of surface functionalisation based on electrochemical hydrogenation followed by electrochemical chlorination of glassy carbon was developed. The so prepared surfaces were used for immobilisation of thiolated molecules including thiolated DNA. Finally the synthesised molecules and modified surfaces were used for electrochemical detection of genetic mutation following the APEX protocol.

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