Hi, everyone. I am performing an ARMS Q-PCR for KRAS mutation detection and using the taqman probe. I wonder if the probe need to follow the ARMS primer or not? The probe I use now is following the non-arms-primer and I find the WT KRAS has also been amplified. Usually, the primers I designed are not quite specific and WT can be amplified but in this case, the CT value of WT KRAS is too small (CT value of WT: 33, CT value of mutation: 29). By the way, the probe I use includes two LNA modified bases. I thought maybe the LNA modified probe was the key of the problem so I tried probe of lower concentration but the result wasn't much different...Now I don't know how to adjust, pls give me some advice.
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