A real-time PCR instrumentation platform consists of a thermal cycler, optics for both fluorescence excitation and emission collection, together with a computer and software for data acquisition and analysis.
A wide range of systems are available and these differ in design and level of sophistication. The variation of features includes: format, reaction vessels, emission and excitation wavelengths, throughput, level of control, chemistry, software, speed and applications. All real-time PCR machines have in common the ability to measure the accumulation of PCR product during the exponential phase of the reaction using online fluorescence monitoring, whether specific or non-specific and hence provide accurate data on initial starting copy numbers.
Amplification and detection are combined in a single step, therefore the process can occur in a single closed reaction vessel eliminating any need for post-PCR manual manipulations, and reducing the possibility of introducing contamination or variability.
Additional technical advantages include both qualitative and quantitative PCR, mutation analysis, multiplexing and high-throughput analysis. Although the fluorescence chemistries used in different platforms are similar, their mechanics and methodologies are wide ranging.from
Logan and Edwards (2009) in Real-Time PCR: Current Technology and Applications
- Real-Time PCR: Current Technology and Applications
- Real-Time PCR in Microbiology: From Diagnosis to Characterization
- PCR Troubleshooting: The Essential Guide
- PCR Books
Labels: PCR instrumentation, qPCR thermal cycler, real time PCR machine