I just want to confirm that the probe in a TaqMan Assay should have a Tm 5-10'C higher than the Forward and Reverse primers and NOT 5-10'C lower? Given a final concentration of 500nM of primers and 250nM of probe and standard salt concentrations etc.....
Yep, that is correct.
To confirm, can check out the PremierBiosoft and Life Technologies overview pages.
http://www.premierbiosoft.com/tech_notes/TaqMan.html
http://www.lifetechnologies.com/us/en/home/references/ambion-tech-support/rtpcr-analysis/general-articles/top-ten-pitfalls-in-quantitative-real-time-pcr-primer.html
Best Regards,
Sergey Kachur
Yep, that is correct.
To confirm, can check out the PremierBiosoft and Life Technologies overview pages.
http://www.premierbiosoft.com/tech_notes/TaqMan.html
http://www.lifetechnologies.com/us/en/home/references/ambion-tech-support/rtpcr-analysis/general-articles/top-ten-pitfalls-in-quantitative-real-time-pcr-primer.html
Best Regards,
Sergey Kachur
Thank you, wanted to confirm that this was the case. You know how it is, you can sometimes forget the basics when you are dealing with a multitude of new techniques and methods.
- Badges
- Science method