Real-Time Polymerase Chain Reaction For Detection Of Optimal Thermal Profile For DNA Templates From Air-Dried Insect Collection Material

The thermal profile for polymerase chain reaction (PCR) technique is the main concern in obtaining the PCR product. Determination of optimal thermal profile can be achieved by various techniques, the main objectives of which are simplicity and cost effectiveness of the method. Real-time PCR is a novel molecular biology method widely used in different fields, recognised by its accuracy and precision. In the present work, the thermal profile was estimated for DNA from air-dried collection material of Notiophilus genus and new sequences were obtained for Gene-Bank. Air-dried entomological collections exist around the world and the use of DNA from such type of material is widespread and offers various unique preferences. The purpose of the study was to assess the suitability of different primer annealing temperatures (42 °C – 55 °C) for universal mitochondrial DNA COI gene primer pair HCO/LCO using real-time PCR method. During this study we demonstrate that real-time PCR could be useful to determine the optimal thermal profile for low-yield DNA for increasing the number of qualitative DNA sequences.