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Accepted (AM)Open Access (License Unspecified), Open
Journal article
A Multifunctional Reactor with Dry-Stored Reagents for Enzymatic Amplification of Nucleic Acids
Analytical chemistry (Washington), v 90(2), pp 1209-1216
16 Jan 2018
PMID: 29226671
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
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.
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Details
- Title
- A Multifunctional Reactor with Dry-Stored Reagents for Enzymatic Amplification of Nucleic Acids
- Creators
- Jinzhao Song - University of PennsylvaniaChangchun Liu - University of PennsylvaniaMichael G. Mauk - Univ Penn, Sch Engn & Appl Sci, Dept Mech Engn & Appl Mech, Philadelphia, PA 19104 USAJing Peng - University of PennsylvaniaThomas Schoenfeld - Lucigen CorporationHaim H. Bau - University of Pennsylvania
- Publication Details
- Analytical chemistry (Washington), v 90(2), pp 1209-1216
- Publisher
- American Chemical Society; Washington, DC
- Number of pages
- 8
- Grant note
- R01CA214072 / NATIONAL CANCER INSTITUTE; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Cancer Institute (NCI) R21DE026700 / NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Dental & Craniofacial Research (NIDCR) 1R43DE025466-01 / NIH; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA 1R21DE026700-01 / NIH NIDCR; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Dental & Craniofacial Research (NIDCR) R01CA214072 / NIH NCI; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Cancer Institute (NCI) R21AI128059 / NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Allergy & Infectious Diseases (NIAID)
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Engineering Technology
- Web of Science ID
- WOS:000423011600026
- Scopus ID
- 2-s2.0-85040671888
- Other Identifier
- 991020623911304721
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- Web of Science research areas
- Chemistry, Analytical