Principle and Purpose
Quantitative PCR (real-time PCR, QPCR) uses the basic principle of classical PCR (cyclic amplification of a DNA fragment based on an enzymological reaction) with the difference that an amplification measured not in final but throughout of the reaction, and therefore in real time.
At each amplification cycle, the amount of DNA is measured using a fluorescent marker such as “syber green” whose emission is directly proportional to the amount of amplicons produced. This makes it possible to obtain a kinetics of the reaction and thus the quantification of the DNA whereas the conventional PCR only gives the final measurement. Thus this technique makes it possible to compare the differences in expression of the genes as functions of the experimental parameters defined by the researcher.
This equipment is completed by a TECAN brand “freedom evo” pipetting robot, which makes it possible to prepare the 96-well PCR plates in record time (around 7 minutes) while avoiding errors of pipetting or contaminations related to user. This pipetting robot is particularly interesting if the number of plates to perform is important. This robot is controlled via TECAN’s Evoware software.
Scientific Support : Vincent LELIEVRE, PR2 UdS