Real time (CT IN NEGETIVE WELL AND DOBLE MELTING CURVE) - (Sep/02/2013 )
i have working on rbohD , after drought stress i cheked the exporession of this gene in my plant.
i have two problem
1. I got High CT in my negetive husekeeping gene (GAPDH) but after check in agarose i didnt see any thing. I am worry about my final result after normalosation with this house keeping gene.
2. sometimes double melting curve in my specific gene only in one accession.
iman_r41 on Mon Sep 2 10:23:07 2013 said:
To point 1. How high was your Ct ? If the Ct is more than 6 cycles higher than the sample with the lowest expression, it should be fine. You are aware that higher Ct stands for lower expression, I assume ?
To point 2 what do you mean by accession ? How does this look on the gel ? Primer dimers perhaps ?