A simple and sensitive technique was developed for the detection of bacterial cells in a water sample using the nested Polymerase Chain Reaction (nested PCR). The test organism, Escherichia coli, was detectable at an absolute value of 1-5 cells/50 mL of spiked water using ethidium bromide staining and ultraviolet light visualization of PCR product formation on DNA agarose gels. Different parameters of the PCR were examined to determine which ones are the most critical in the design of a detection method for very low numbers of cells in potable water samples. The presence of the filters used for sample concentration in the PCR reaction tube was found to be the most inhibitory component of the procedure and the nested PCR protocol was used to circumvent this inhibitory effect.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.22745 |
| Date | January 1994 |
| Creators | Juck, David F. |
| Contributors | Ingram, J. M. (advisor) |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Master of Science (Department of Natural Resource Sciences.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 001462249, proquestno: MM05568, Theses scanned by UMI/ProQuest. |
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