Inexplicable qPCR failure in single wells - (Aug/18/2013 )

No, unfortunately I didn't. It didn't stop either, so I just had to learn to live with it (i.e. repeat the experiment when it happened).

Hi!! 

I was searching in the internet because I' having this kind of problem in my qPCR experiments, and I have just found this discussion. I would like to know if you have already solved the problem and what it was??

Which kind of PCR machine are you using?

I am using a BioRad CFX384. Since I start with my experiments, I always get some wells in which non signal detection is found. As in your case, it is always in one or two of the 3 replicates I use, but the other values are normally OK. I also realized that the electronic pipet I was using to pipet the mix for the replicates in the three wells was not working properly. This can explain a deviation in the Cq value, but I think it does not explain "non-detection", specially when for the other replicates I get a good Cq value...

I have also thought it is the machine, although as in your case, I don't get the errors always in the same wells but, it cannot be explain due to a pipetting error. 

By the moment we will try to check somehow if the machine is working well, because none else is working with it now, so I cannot check if other people is having the same problem. Normally the brands can send kind of a standard plate and then you can send them the results you get with this plate, so like this you know if the machine calibration is Ok. I will try to do this to be sure about this and if not...keep on looking for the thing that is causing the error.

Hi noetm,

I didn't find the problem - the situation is unchanged from my last post but I also didn't follow it up any more because I don't work there any more. In my case it was a Applied Biosystems StepOnePlus machine. Wish you good luck in resolving the issue !! If you find out the problem, it would be great if you could post it here.