January 1, 2013.
Propidium monoazide (PMA) is a molecular tool used to assess viability of microorganisms. Currently, PMA is thought to discern viability through membrane permeability; PMA enters only membrane compromised cells, irreversibly crosslinks to theirDNAand precipitates theDNAout of solution, preventing it from being amplified during polymerase chain reaction (PCR). Using PMA on a sample of live and dead microorganisms results in only theDNAof living organisms being amplified and identified. Therefore, a comparison ofPCRresults with and without PMA allows one to determine the live fraction and total population, respectively.
Current literature provides conflicting evidence as to the effectiveness of the technique. Our research hopes to resolve these differences. Initial results have shown that the length of the targeted section affects results of combined PMA-qPCR. The current model does not account for why this difference might occur, and has led us to conclude that the current model is incomplete and that PMA could also inhibit the binding and/or processivity of enzymes interacting with PMA treatedDNA.
Biology | Cell Biology | Molecular Biology | Pathogenic Microbiology
Nicholas Fingland and Adrian Ponce
NASA Jet Propulsion Laboratory (JPL)
This material is based upon work supported by the S.D. Bechtel, Jr. Foundation and by the National Science Foundation under Grant No. 0952013. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of the S.D. Bechtel, Jr. Foundation or the National Science Foundation. This project has also been made possible with support of the National Marine Sanctuary Foundation. The STAR program is administered by the Cal Poly Center for Excellence in Science and Mathematics Education (CESaME) on behalf of the California State University (CSU).