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Laboratory q-PCR and Culture Techniques

Course Numbers: OCMB - 6001, MEVS - 6001, and CZMT - 6001

The first week of the course will be training in the classical culture techniques for determination of fecal pollution in surface waters as is used in all water quality laboratories. This will involve membrane filtration for bacterial indicators and plaque formation for viral indicators. 

During the second week newly developed real time PCR methods will be performed and evaluated.
The use of q-PCR has been implemented in microbiology studies to quantify abundance and expression of taxonomic and functional gene markers that pose contamination threats to drinking, recreational, marine, and fresh waters. Its use allows viable results for the indication of microbial presence associated with human pollution that supersedes the abilities of culture based fecal coliform and enterococci studies. The use of PCR chemistries is a more advanced, precise and sensitive method for estimating microbial species in environments. Within PCR chemistries, q-PCR allows for expedient results coupled with greater accuracy to determine if human pollution is contaminating a water source and in what amounts quantitatively.

Learning Objectives

Students will:

  • Understand the significance of indicator microorganism to fecal water pollution studies
  • Become familiar with classical culture procedures for determination of fecal coliform and enterococci and the interpretation of their significance
  • Be able to to identify parameters necessary to complete basic stock assessment modeling problems
  • Become familiar with q-PCR procedures to determine the same indicator species as done with culture methods
  • Establish advantages and disadvantages of both techniques and relative specificity to human waste

Textbook:

No textbook required. Posted citations available online. 

Winter 2016 Syllabus

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