Design and development of an electronic instrumentation device for high-resolution melting curve analysis

Identifier:  TFM:186

Handle: https://hdl.handle.net/20.500.11797/TFM186

Authors:  Escobar Nassar, Santiago

Abstract:
Abstract. The design and development of a computer-controlled heating block is reported. This unit is part of a novel electronic instrumentation system, specially designed for electrochemically marked high resolution melting curve analysis and intended to be used for the multiplexed detection of SNPs and mutations. The system herein explained, consists of a pair of aluminum blocks that are heated in a controlled way using a pulse width modulation protocol implemented on an Arduino UNO. Data processing and temperature control at different heating resolutions was performed via a proportional integral controller, also implemented on the Arduino. Heating tests using as template a glass slide with PMMA (setup that simulates the behavior of real electrodes) were performed to validate the system functionality and performance. Furthermore, a 1 and 13 gold electrode arrays were modified with a double thiolated covalently linked polyT single stranded DNA. Afterwards, the system was incubated with a polyA chain labelled with horseradish peroxidase (HRP). This system was exposed to a tetramethylbenzidine dihydrochloride (TMB) and hydrogen peroxide (H2O2) solution to determine the best electrode coverage ratio. A fast chonoamperometric test was carried out to determine the best probe/backfiller ratio for forthcoming experimentations on a model DNA sample