Does anybody use 7500 Real time PCR of Applied Biosystems. I want to use touchdown PCR setting but don't know how to set. Can anybody help me?
Does it have a option for insertion of individual steps in programming.?. If yes, then you can make a step down PCR.
Follow the link and see "How do I set up a touchdown PCR experiment?"
http://www.lifetechnologies.com/us/en/home/technical-resources/technical-reference-library/real-time-digital-PCR-instruments-support-center/7500-real-time-pcr-systems-support/7500-real-time-pcr-systems-support-getting-started.html
Dear Saravana,
I also checked the attachment before, but it looks completely different from the 7500 PCR machine that we have.
Is a touchdown protocol really compatible with a qPCR measurement?? It seems like in the early (high anneal temp) rounds of a touchdown PCR, you are likely sacrificing efficiency to achieve specificity. And the mathematical basis of qPCR quantitation assumes that a) PCR efficiency is the same throughout all of the cycling steps--especially the early ones and b) that the PCR efficiency of the gene of interest assay is the same as the PCR efficiency of the reference gene. So you might be able to rank the quantities in samples using touchdown qPCR, but I don't think that the usual math will work.
I also have the same question as David F. berker. Can we really quantify the template with touchdown PCR?
- Badges
- Science topic