Real-Time PCR: Current Technology and Applications is "a useful compendium of basic and advanced applications for laboratory scientists. It is an ideal introductory textbook and will serve as a practical handbook in laboratories where the technology is employed."

from Christopher J. McIver, Microbiology Department, Prince of Wales Hospital, New South Wales, Australia in Australian J. Med. Sci. 2009. 30(2): 59-60

Further reading: Real-Time PCR: Current Technology and Applications

Labels: book review, books, PCR, qPCR

Now available on the web: A comparison of the following PCR machines lists various features to help you decide which qPCR instrument is most suitable for your needs.

• Applied Biosystems: ABI 7300, ABI 7500, ABI 7500 Fast, ABI 7900 Fast HT with automation accessory, ABI StepOne
• Roche: LightCycler 480, LightCycler 1.5, LightCycler 2.0
• Stratagene: Mx4000, Mx3000P, Mx3005P
• Cepheid: SmartCycler
• Corbett: Rotor-Gene 6000
• Eppendorf: Mastercycler ep realplex
• BioRad: MiniOpticon, MyiQ, Opticon2, Chromo4, iQ5

read more ...

Bibliography:

1. Real-Time PCR: Current Technology and Applications
2. Real-Time PCR in Microbiology: From Diagnosis to Characterization
3. PCR Troubleshooting: The Essential Guide
4. PCR Books

Labels: instruments, molecular machines, platforms, qPCR, real-time PCR

In weighing up the pros and cons of the different real-time PCR machines for your laboratory, factors to consider include: supported chemistries; multiplex capability for that chemistry; throughput; flexibility; format; easy-of-use and robust software package; reproducibility; speed; size; technical support; customer support and not least the cost, not only of the initial equipment outlay and servicing but also the associated cost of consumables and reagents. It is also possible to 'try before you buy', most companies will provide a loan machine. It is wise to test a few of these once you have narrowed down your choice. User experiences should not be overlooked and there are now a number of useful websites and news groups where you can address you questions and queries regarding PCR machines read more ...

Labels: PCR, PCR machine, qPCR, qPCR machine, qPCR platform

November 16 - 18, 2009 qPCR Congress
London, UK Further information
The advancement of real-time or quantitative polymerase chain reaction (qPCR) extends the usefulness of PCR technology. This conference explores effective strategies, technologies and applications of qPCR in accelerating drug discovery and development of clinical diagnostics.
Suggested reading: qPCR Books

Further reading: Real-Time PCR in Microbiology: From Diagnosis to Characterization

Labels: qPCR, qPCR conference

Real-time PCR has removed many of the limitations of standard end-point PCR and since its introduction in the mid-1990s there has been an explosion both in the number of publications and available instrumentation describing real-time PCR applications across many disciplines. Real-time PCR (RT-PCR) technology is highly flexible and many alternative instruments and fluorescent probe systems have been developed recently. The decreased hands-on time, increased reliability and improved quantitative accuracy of RT-PCR methods have contributed to the adoption of RT-PCR for a wide range of new applications.

The development of instruments that allowed real-time monitoring of fluorescence within PCR reaction vessels was a significant advance. The technology is flexible and many alternative instruments and fluorescent probe systems are available. RT-PCR assays can be completed rapidly since no manipulations are required after the amplification. Identification of the amplification products by probe detection in real-time is highly accurate compared with the traditional PCR method of size analysis on gels. Analysis of the progress of the reaction allows accurate quantification of the target sequence over a very wide dynamic range, provided suitable standards are available. Further investigation of the RT-PCR products within the original reaction mixture using probes and melting analysis can detect sequence variants including single base mutations. RT-PCR has found applications in many branches of biological science. Applications include gene expression analysis, the diagnosis of infectious disease and human genetic testing. Due to their fluorimetry capabilities, these real-time machines are also compatible with alternative amplification methods such as NASBA, provided a fluorescence end-point is available.

The introduction of RT-PCR assays to the clinical microbiology laboratory has led to significant improvements in the diagnosis of infectious disease. The technology has applications in clinical bacteriology, parasitology and virology. There are few areas of clinical microbiology which remain unaffected by this new method. It has been particularly useful to detect slow growing or difficult to grow infectious agents.Its greatest impact is probably its use for the quantitation of target organisms in samples. The ability to monitor the PCR reaction in real-time allows accurate quantitation of target sequence over at least six orders of magnitude. The closed-tube format which removes the need for post-amplification manipulation of the PCR products also reduces the likelihood of amplicon carryover to subsequent reactions reducing the risk of false-positives. Standardisation of assay protocols for use in diagnostic clinical microbiology and external quality control schemes is required to ensure quality of testing.

Further reading:
Real-Time PCR: Current Technology and Applications
Real-Time PCR in Microbiology: From Diagnosis to Characterization

Labels: diagnostics, PCR, qPCR, real-time PCR, RT-PCR