Bad primer efficiency during multiplex qPCR - (May/29/2008 )

Hello,

I am doing qRT-PCR on leukemia samples looking at two genes (A and B) using Taqman. One of my primers (B) does not appear to be working well with my internal standard © human PO (E=1.7-1.9). I am now looking at the primers separately usng a standard curve to verify that the efficiency of my primers of interest (A and B) do change in the presence of the internal standard primers. I have done the following set-up: A, B, C, AC, BC. In case this doesn't work does anyone have any suggestions as to an additional internal standards I can use for leukemia samples? BTW I cannot use B-actin, it is known to be diff. expressed in leukemia.

Hello,

I am doing qRT-PCR on leukemia samples looking at two genes (A and using Taqman. One of my primers ( does not appear to be working well with my internal standard © human PO (E=1.7-1.9). I am now looking at the primers separately usng a standard curve to verify that the efficiency of my primers of interest (A and do change in the presence of the internal standard primers. I have done the following set-up: A, B, C, AC, BC. In case this doesn't work does anyone have any suggestions as to an additional internal standards I can use for leukemia samples? BTW I cannot use B-actin, it is known to be diff. expressed in leukemia.