Techne PrimeQ Real time PCR Reaction Volume
When using Techne PrimeQ we recommend a sample volume of 20µl, which in itself is economical in the use of costly reagents. The application note A08-001A, which can be found online demonstrates how this can be reduced by half to 10µl with no loss of reaction efficiency and little effect on sensitivity. Final fluorescence levels were reduced but this was to be expected due to the smaller amount of dye available in the mix. The use of low profile plates in the Techne PrimeQ is also of benefit since there is reduced distance between the sample and detection mechanism.
Volume reduction is therefore applicable for robust PCR assays where copy numbers are reasonably high and sensitivity is not an issue. We would not however recommend reducing the sample volume significantly for sensitive assays.
Contact camlab for more information on the Techne PrimeQ Real time PCR system