Protocol run times (e.g., for fast PCR).Whether gradient capability is required.Trade-offs between yield and specificity.The range of sample volumes (analytical to preparative).The number of samples to be amplified at one time.Considerations when choosing a thermocycler include the following: Which thermocycler, or thermocyclers, you choose depends on current and anticipated future needs. There is an increasing array of types and uses of polymerase chain reaction, from reverse transcriptase PCR (RT-PCR) to Droplet Digital™ PCR. More recently, PCR assays and the ability to fully integrate a thermocycler into automated workflow have made pathogen testing faster and more sensitive, with a higher throughput. Validation studies have shown that the PCR kits are equivalent to, or better than, the various reference methods evaluated while providing results in much less time.Ĭulturing and ELISA were the main assays for testing for major food pathogens for many years.
Walk-away automation provides automated DNA extraction and PCR setup, combined with automated data collection and analysis, bar codes for increased traceability, and LIMS integration.įurther, PCR kits are validated by several certification bodies. Use of automated PCR for pathogen testing, in addition to increasing speed, provides a higher throughput, lowers contamination, reduces the level of false positives, and decreases interlab variability.
This shortens the overall time to getting a result and minimizes required hands-on time. Increased sensitivity means that an enrichment step is less likely to be required prior to assay. PCR is both more accurate and rapid than ELISA. This pathogen can be detected using a PCR kit in as little as 12 hours. coli O157:H7, speed and sensitivity is critical. Many foodstuffs are perishable, so testing has to be fairly rapid or the food will not have a sufficient shelf life after testing is completed. Speed is an important consideration for testing foods for pathogens. Thermocycler Use in Food Pathogen Detection A number of fun PCR amplification kits are available for teaching real-world PCR applications, for example, the Crime Scene Investigator PCR Basics™ kit and the Fish DNA Barcoding kit. Students can learn how PCR is used in fields such as forensics, diagnostics, animal conservation, and archaeology. While learning how PCR analysis is used in practical ways, students are exposed to a range of scientific principles.
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Integration of a memorable hands-on approach into the curriculum demonstrates practical applications of science, helps with retaining information, teaches how to interpret data, and keeps students engaged.įor life science education, a thermocycler is a good teaching tool for inquiry-based science. Inquiry-based science teaching helps develop critical thinking skills. Using these PCR machines, students will gain a real research experience and an understanding of how PCR is being used in everyday testing. All thermocyclers are fully functional standard machines. Several PCR machines and real-time PCR systems are available for educational use. The controlled temperature ramping provided by a thermocycler enables the study of temperature-dependent kinetics for example, in addition to optimizing a PCR protocol, a thermocycler with a gradient feature allows for the determination of the optimum temperature for any enzymatic activity. A thermocycler can be used in protocols where very strict temperature control is required such as heating tissue slides for in situ hybridization. The fine temperature control and ability to hold a precisely set temperature with little fluctuation is also applicable to other techniques in which high-accuracy temperature control increases reproducibility between experiments. Thermocycler use extends beyond simple PCR and the amplification of nucleic acids to include both random and site-directed mutagenesis as well as the in vitro construction of recombinant DNA sequences. In molecular biology they are used for DNA sequencing, cloning, generation of probes, quantification of DNA and RNA, studying patterns of gene expression, detection of sequence-tagged sites, and many more techniques. Thermocyclers are integral to life science research. Thermocycler Use in Life Science Research
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