7500 Fast RT PCR machine
The 7500 Fast is a real-time PCR machine which enables detection and quantification of PCR products in real-time using either SYBR green reagents or Taqman probes. It is possible to monitor PCR reactions cycle by cycle enabling quantification and rapid analysis of many different targets.
Most users within the school use SYBR Green reagents which emit a fluorescent signal upon binding to double-stranded DNA molecules. This allows the amount of each PCR product being synthesised to be measured, giving either relative or actual measurements of mRNA (via RT-PCR) or DNA levels. The results are plotted as a sigmoidal curve in which the number of cycles are plotted against fluorescence. The threshold cycle (CT), is used to calculate the starting amounts of template DNA.
This technology can be used in number of applications including measuring gene expression, pathogen detection and in post PCR non quantitative assays for allelic discrimination. Within the School it is being widely used to measure gene expression. If you require further information or training to become an authorised user please contact Liz McCarthy or Rachel Gregory.
Protocols can be found on the Applied Biosystems website
