What feature of real time PCR matters most?
1. Cost of plasticware- plates/tubes
or
2. Cost of reaction mixture- master mix, fluorescent probes
or
3. number of total reaction that can be run simultaneously; considering the different Tm values for different primers
Real-time PCR allows you to quantify your target genes, as well as quickly see the results without running gel electrophoresis! Probe are rather expensive, but the most expensive the QPCR cycler! Note that not all machines have thermal gradient function allowing you to run samples with different primers simultaneously!
You should consider all factors,
You can also design primers with similar Tm so you don't have to run gradients. I have designed dozens of primers this way without an issue.
Don't forget that you can build your own, especially if you have both an engineering dept and a computer dept that will help build it. Search openpcr for hints.
There are at least these:
http://www.instructables.com/id/Arduino-PCR-thermal-cycler-for-under-85/
http://openpcr.org/
http://www.instructables.com/id/Coffee-Cup-PCR-Thermocycler-costing-under-350/step2/Completed-version-of-the-Coffee-Cup-Thermocycler/
Definitely, the cost of reagents matters the most and it is the thing where you will be spending the most of your money. Purchase some open ended system like ABI machines. You can then step wise decrease the cost of the reagents (master mix, assay mix, etc.) by your experience and moving towards multiplexing.
- Badges
- Science topic