Real-time PCR Chemistry: Choice of Platform

Second, third and fourth generation real-time PCR platforms have been developed with improvements in multiplexing and increased throughput capabilities. The optical characteristics of a given platform clearly have an impact on the ability to multiplex and also determine which probe systems are compatible. In addition, the analysis software may also predetermine the appropriate chemistries.

Iit is important to point out that the platform and choice of fluorescent chemistry are strongly linked. Indeed some platforms are biased towards a particular probe system and whilst the optics permit different probe chemistries to be excited and detected, often the analysis software does not and the user is required to export the data to a spreadsheet program for detailed user analysis. For example, the Applied Biosystems platforms do not officially support any chemistries other than hydrolysis probes and SYBR Green and only support duplexing (Logan and Edwards, 2009). Therefore, the reporting chemistry required for an application should be strongly considered before a choice of platform is made.

from Logan and Edwards (2009) in Real-Time PCR: Current Technology and Applications

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: Applied Biosystems, chemistries, chemistry, duplexing, fluorescent chemistry, hydrolysis probes, multiplexing, SYBR Green