I´m designing probes for qPCR multiplex with Taqman chemistry for gene expression.
What they consider should be the ideal size (range) of PCR products?
Thanks a lot for your time.
Rolando.
Between 100 and 200bp is generally pretty optimal for qPCR, but you can go outside that range, I've had good luck with 80-90bp amplicons as well as 250+
Hi Jesus
Craig is right, but since you a multiplex condition and their is a competition between fragments, I would prefer to keep them around 100 bp. Otherwise smaller fragments (100 bp) would have a higher chance of amplification in compare to larger fragments (250 bp +), specially when you have more than 2 fragments.
Best of luck
It depends on the machine and system. For ABI type detection with different flu. chromes, theoretically, doesn't need to be any difference. If you are using one flu chrome, then I suggest a minimum of 25 bp. If you are using a general system (Ethidium Bromide) probable a minimum of 50 bp is recommended by our lab.
Hope this helps.
Bob
- Badges
- Science topic
- Similar topics
- Chemistry