Can you please help me as I made a stupid mistake? I have duplex TaqMan qPCR assay. I purchased FAM & VIC probes for my duplex assay. Unite size was 6000 pmol for each probe. When I received my probes, I've completely forgotten to create stock solutions of 100µM in x1TE. I've seen on the sheet unit size 6000 pmol; volume 60µl. Therefore, I've considered my probes as already prepared stocks of 100 µM. I went from there and prepared my working solutions to be 5 µM (5ul of 6000pmol probe and 95 ul of sterile MilliQ water).
I've stupidly aliquoted 5ul of 6000 pmol of each probe into tubes and stored at -20C.
What to do from here? How can I test if a screwed concentration of probes have impact on final number of copies of target and internal control. I still have aliquots of 5ul of 6000 pmol of each probe in the freezer.
I am half way through project and had November as deadline for results delivery and limited money on the project account.
Your help is much appreciated.