Song, Jinzhao Liu, Changchun G. Mauk, Michael Peng, Jing Schoenfeld, Thomas Bau, Haim H. A Multifunctional Reactor with Dry-Stored Reagents for Enzymatic Amplification of Nucleic Acids To enable inexpensive molecular detection at the point-of-care and at home with minimal or no instrumentation, it is necessary to streamline unit operations and store reagents refrigeration-free. To address this need, a multifunctional enzymatic amplification reactor that combines solid-phase nucleic acid extraction, concentration, and purification; refrigeration-free storage of reagents with just-in-time release; and enzymatic amplification is designed, prototyped, and tested. A nucleic acid isolation membrane is placed at the reactor’s inlet, and paraffin-encapsulated reagents are prestored within the reactor. When a sample mixed with chaotropic agents is filtered through the nucleic acid isolation membrane, the membrane binds nucleic acids from the sample. Importantly, the sample volume is decoupled from the reaction volume, enabling the use of relatively large sample volumes for high sensitivity. When the amplification reactor’s temperature increases to its operating level, the paraffin encapsulating the reagents melts and moves out of the way. The reagents are hydrated, just-in-time, and the polymerase reaction proceeds. The amplification process can be monitored, in real-time. We demonstrate our reactors’ ability to amplify both DNA and RNA targets using polymerase with both reverse-transcriptase and strand displacement activities to obtain sensitivities on-par with benchtop equipment and a shelf life exceeding 6 months. polymerase reaction proceeds;sample volumes;strand displacement activities;paraffin-encapsulated reagents;acid extraction;sensitivity;paraffin encapsulating;shelf life;6 months;sample volume;RNA targets;sensitivities on-par;reaction volume;amplification process;acid isolation membrane;store reagents refrigeration-free;unit operations;DNA;refrigeration-free storage;Dry-Stored Reagents;Nucleic Acids;Multifunctional Reactor;chaotropic agents;benchtop equipment;Enzymatic Amplification;amplification reactor 2017-12-11
    https://acs.figshare.com/articles/media/A_Multifunctional_Reactor_with_Dry-Stored_Reagents_for_Enzymatic_Amplification_of_Nucleic_Acids/5729451
10.1021/acs.analchem.7b03834.s001