Multiplex Quantitative MiRNA Analysis On A Real Time Slipchip System

PCR is a technique widely used in research and diagnostics for nucleic acid quantification.Measuring the concentration of different RNA and DNA targets provide useful information related to medical conditions,infections or diseases.Several approaches can be addressed for this task,such as Digital PCR(d PCR)and Real-Time or Quantitative PCR(qPCR).While d PCR performs an end-point absolute quantification,qPCR allow a broader multiplex analysis.This advantage allows fast quantification of several target within a same sample,reducing experiments’ times and costs.Nevertheless,current qPCR systems are either bulky,expensive or with low multiplexity availability.Under these premises,the use of microfluidic technology can open the path for future qPCR systems.In this thesis,a qPCR system based on Slip Chip technology has been proposed and developed.Slip Chip is a microfluidic approach that consists in the relative movement of two plates in which microwells and channels are carved.One position allows easy loading of a sample in the microwells without the need of valves or pumps,and the slipping movement of both plates isolates the sample in to up of thousands of microwells,making it ideal for multiplexing.Based on this technology,as Raspberry Pi-based system has been proposed in which the PCR process is carried out within the Slip Chip and controlled by the developed system.Moreover,it will be constrained within a compact,cost-efficient design.