OWML: QA/QC

Quality Assurance/Quality Control Manual: Ohio Water Microbiology Laboratory

Thermal Cyclers
There are two thermal cyclers used to perform the polymerase chain reaction (PCR). The PCR is done to amplify microbial DNA targets through a series of temperatures changes. Numerous projects use the PCR for a variety of applications including microbial source tracking, detection and/or quantification of enteric viruses, and detection and/or quantification of bacterial indicators. The thermal cycler used for end-point PCR performs the PCR to completion before the amplified DNA products can be subsequently detected using gel electrophoresis. The thermal cycler used for real-time quantitative PCR (qPCR) detects and quantifies the targeted DNA sequence as it is being amplified.

End-point PCR thermal cycler

• The temperature is monitored twice a year.
• Results of the temperature QC are kept in the Master Cycler QC logbook.
• Temperature monitoring dates are recorded in the LIMS.
   

qPCR thermal cycler

• Background calibration is performed monthly.
• A pure-spectra assay is done annually or as needed (if replicate amplification curves are continually wavy or non-uniform).
• A RNase P verification run is done as needed (as a troubleshooting measure).
• A regions of interest (ROI) plate is run as needed (following lamp replacement or if the system is jostled).
• All quality assurance checks are recorded in the LIMS.