The majority of instrument manufacturers supply optimised real-time PCR mastermixes, these reagents benefit from being quality controlled, are easy to use, and usually offer reproducible and reliable results. However, for certain laboratories the additional cost can be prohibitive, however with an array of third party companies (e.g. Qiagen, Eurogentec, Invitrogen) now supplying real-time reagents, competition in this market should lead to reduced costs. Target specific kits for a range of applications are available from some manufacturers and other companies (e.g. Roche, Applied Biosystems, Qiagen). Additionally, Applied Biosystems offer Assays-on-Demand and Assays-by-Design services for SNP genotyping by real-time PCR. A number of companies now offer real-time PCR services from primer design right through to assay design and validation, quality assurance and other custom services.
from Logan and Edwards (2009)
in Real-Time PCR: Current Technology and ApplicationsBibliography:
- Real-Time PCR: Current Technology and Applications
- Real-Time PCR in Microbiology: From Diagnosis to Characterization
- PCR Troubleshooting: The Essential Guide
- PCR Books
Labels: Applied Biosystems, Eurogentec, Invitrogen, PCR mastermixes, Qiagen
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 ApplicationsBibliography:
- Real-Time PCR: Current Technology and Applications
- Real-Time PCR in Microbiology: From Diagnosis to Characterization
- PCR Troubleshooting: The Essential Guide
- PCR Books
Labels: Applied Biosystems, chemistries, chemistry, duplexing, fluorescent chemistry, hydrolysis probes, multiplexing, SYBR Green