NPSA for Specific and Dye-Based Subattomolar Nucleic Acid Detection at 37 °C
We describe a nested phosphorothioated (PS) hybrid primer-mediated isothermal amplification (NPSA) assay which only utilizes EvaGreen (a DNA-binding dye) to achieve specific and dye-based subattomolar nucleic acid detection at 37 °C. The success of low-temperature NPSA essentially depends on employing Bacillus smithii DNA polymerase, a strand-displacing DNA polymerase with wide range of activation temperature. However, the NPSA’s … More NPSA for Specific and Dye-Based Subattomolar Nucleic Acid Detection at 37 °C