TaqMan probe - (Feb/13/2006 )

Can anyone tell me, what's the standard concentration of TaqMan probe used for Real Time PCR. Because I'm working with a final concentration of 1 µM and I don't know if that's normal.

Thanks

Nieky

Nieky, 1 µM is incredibly high. The maximum amount I have ever seen recommended by any company (ABGene, ABI, etc.) is 250 nM probe concentration. What are your results like?

Thanks for the respons.
My results are strange. I was thinking, could it be, because my probe begins with a G and that this G is quenching the reporter dye?


Nieky

I´ve got a different problem with taqman probes.
I´ve set up the machine to read at the extension step and it´s reading a very high relative fluorescence at the very beginning of the PCR. I wonder if this is the step when I have to read
the fluorescence with a taqman probe reaction.
Thanks in advance,

Pablo

we've set the detection of the fluorescense during the annealing step. the first time I also did it during extention but it didn't gave any results. with the detection during annealing you will get results!

good luck

Nieky