PMC:7143804 / 38512-39489 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/7143804","sourcedb":"PMC","sourceid":"7143804","source_url":"https://www.ncbi.nlm.nih.gov/pmc/7143804","text":"First, to determine the temperature window of operation, MDA reactions are performed at 25 °C and 30 °C using the Illustra GenomiPhi V2 DNA amplification kit and EvaGreen fluorescence dye. From the literature, we know that this reaction does not work above 35 °C due to degradation of the protein activity in presence of a substrate [44]. In Figure 13, a graph of the fluorescence signal during MDA reactions at 25 °C and 30 °C, together with their non template control (NTC) is shown. These reactions are carried out in a conventional Bio-Rad CFX96 Touch Real-Time PCR machine (Bio-Rad Laboratories, Inc., Hercules, CA, USA) and the results show that the chosen proof-of-principle DNA amplification reaction is temperature dependent to some extent, but that there is a wide range of temperatures at which the amplification can be performed, i.e., 25°C to 35°C. This makes the functioning of the integrated resistive heater less critical than the stability shown in Figure 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