New Approach of PCR Technology for IGF-I Evaluation

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New Approach of PCR Technology for IGF-I Evaluation

Journal Title: Biomedical Journal of Scientific & Technical Research (BJSTR) - Year 2018, Vol 8, Issue 4

Abstract

PCR or Polymerase Chain Reaction is a technique widely used in molecular biology laboratories. Since it was developed in 1983, many procedures use this technique as a basic principle, for example: forensic tests, diagnosis of diseases, phylogenetic studies, identification of mutations, nucleotide sequencing, among others [1-3]. In our procedures of IGF-I diagnostic, we use the techniques of northern blot, western blot, Elisa, immunocytochemistry and cytofluorometry, and the technique of PCR seems still the most rapid and useful in clinical laboratory [4,5] The IGF-I test is among the most important indications for precancerous diagnostic [6]. Therefore, the accessibility of PCR technique giving increased efficiency and rapidity has become a goal to establish a new methodology. The traditional thermal cycler consists of a metal plate or block that has the Eppendorf tubes located at the top, and at the bottom is an electrical device that uses Peltier technology to increase or decrease the temperature according to the requirements of the cycle. There are temperature sensors distributed below the block (Figure 1). For PCR tests, a commercial thermal cycler is relatively expensive. There are several models of low cost thermal cyclers, but they are generally inefficient or inaccurate and also have high energy consumption which is an obstacle for mobile laboratories supported by a limited energy source [7]. Given this challenge, we have developed a low-cost thermal cycler that offers better efficiency with lower energy consumption, an important asset to enable the use of PCR technique in developing countries. Methodology This thermal cycler was designed with the purpose of decreasing the time it takes the sample to reach the desired temperatures. To achieve this goal, it was necessary to change the principle of operation of the machine; instead of using chillers and heaters, the machine uses an Arduino board to control a mechanical system composed of two motors, two conveyor belts, two transverse pinions and a tray where the samples are located (Figure 2) [8]. This system introduces and removes the samples in a bath of glycerin hot at 108 °C, the program of the microcontroller constantly monitors the temperature inside the Eppendorf tube, in this way the exact time is used and necessary to complete the reaction.

Authors and Affiliations

Ricardo Santander, Jerzy Trojan

Keywords

Polymerase chain reaction Forensic tests Phylogenetic studies Immunocytochemistry Cytofluorometry Precancerous diagnostic Eppendorf tube Arduino board Peltier device Conventional thermal cycler

EP ID EP588535
DOI 10.26717/BJSTR.2018.08.001678
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