qPCR machines. Part 3
Ideally, the analysis software supplied with the platform should be as user-friendly as possible but it is also important to check that the software can fully analyse results of the chosen probe chemistry. Some platforms and analysis software suites are biased towards certain chemistries. Some real-time instruments also have specific primer and probe design software that is either supplied with the hardware or available at extra cost. Such software can help simplify and speed up the assay design process and is optimised for that system and reagents. The LightCyclers also have specific relative quantification software that is designed to determine the exact relative nucleic acid concentration normalized to a calibrator sample. This software speeds up and greatly simplifies relative quantification.
from Logan and Edwards (2009) in Real-Time PCR: Current Technology and Applications
Bibliography:
from Logan and Edwards (2009) in Real-Time PCR: Current Technology and Applications
Bibliography:
- Real-Time PCR: Current Technology and Applications
- Real-Time PCR in Microbiology: From Diagnosis to Characterization
- PCR Troubleshooting: The Essential Guide
- PCR Books
Labels: qPCR, qPCR software, qPCR thermal cycler, qRT-PCR, RT-PCR software, software
Real-time PCR platforms
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
Bibliography:
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
Bibliography:
- 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
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