qPCR - Calibration test of Opticon 2

Due to results of Opticon testing from 20090722, we have acquired FAM Calibartion Dye from Bio-Rad. Although not listed online or on the product itself, Bio-Rad customer service informed me that the concentration = 1mM. The calibration protocol in the Opticon 2 manual (p. 10-4) says to use 0.3uM (Cf) in 50uL. Will follow this info. Made up 15mLs of dye solution and distributed 50uL into each well of 3 plates. Tested all three plates on two different machines (Opticon 2 and Friedman lab’s).

Results: This did not produce any useable information whatsoever. In fact, the Friedman lab’s machine didn’t detect any signal at all from any of the three plates. Not sure what the story is.

UPDATE Received email correspondence from Carl Fisher of Bio-Rad and he has indicated that this is not the proper dye and that the conentration is not anywhere near 1mM (although he can’t find any info on what the concentration might be). He has told me to order a different part number (10006046) that is NOT listed anywhere on the Bio-Rad website.