Real-Time PCR Question - (Nov/08/2006 )
hello-
what does it mean when you amplify two mRNAs and one amplification curve has a low ct but the other curve (with a higher cT) has a much higher end point fluorescence reading?
could amplicon size be a factor?
-BioDude1-
QUOTE (BioDude1 @ Nov 8 2006, 05:56 PM)
hello-
what does it mean when you amplify two mRNAs and one amplification curve has a low ct but the other curve (with a higher cT) has a much higher end point fluorescence reading?
could amplicon size be a factor?
what does it mean when you amplify two mRNAs and one amplification curve has a low ct but the other curve (with a higher cT) has a much higher end point fluorescence reading?
could amplicon size be a factor?
Are you using SYBR green?
My guess is that you are having issues around non-specific products amplifying. SYBR green will detect these just like it detects "correct" product, and give you an erroneous idea of product.
You can run the end product on a gel to check, or use the dissociation step.
More details would be helpful.
-Patty4150-