Entering edit mode
7.7 years ago
mer.tantawy
•
0
Dear All
hello, my question is in my data exported from qRT-PCR I have Ct more than 35, or undetermined in miRNA profiling. when I detect the mean of them how to put the ct more than 35 or undetermined, change it to 35 or remove it and make it blank.
thanks
Marwa
You could put the number 90 because that is more than 35. The right course of action is determined by your goals. It seems like the miRNa is not amplifying, simply not detected. Could be zero quantity or a bad reagent. This is not a math question.
You need to provide more detail. Usually you want below 35CT cycles. You could maybe optimize your primer amounts if you haven't done so.
Other explanations:
Not much of that miRNA transcript in your starting template? Primer specificity may be off? Redesign primers ( additionally redesign around functional domain regions) Your working stock dilutions are too diluted? Try working with more concentrated DNA?
What are your primer efficiencies like?