Posted 17 September 2009 - 05:16 PM
My no template control had a positive result. which was strange considering I used water. Some of the other controls that should have been negative had Cp values. They were high though. Also the fluorescence was around 0.31 to 0.5 for those that should be negative.
my positive control did not work. I think the cDNA degraded. I have had previous success using the cDNA from the same sample where it had a Cp value of around 21.
My questions are does anyone know how to determine positive or negative results using Cp values. I have read some research articles where they say if the results are <35 or less than <38 then its negative. How do they come up with these values and justify it for publication?
The stuff I am working on I am hoping to publish but I cannot have shady data.
Posted 17 September 2009 - 05:18 PM
Does anyone know if you can upload positive control data from a previous experiment using the LightCycler software?
Posted 18 September 2009 - 12:59 AM
The first thing comes to mind is that you have a pcr-product contamination.