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High initial fluorescence - need help with some results... (Oct/18/2007 )

Hi!! Can you help me with this? wacko.gif

This is the situation:

- a particular cDNA, 2 10-fold dilution (1 and 2)
- dilution 2 shows initial fluorescence (Rn) at 1...
- dilution 1 shows initial fluorescence (Rn) at 3!!

So when the program (MxPro) plots the fluorescence as dRn, both dilution show similar Cts... = BAD RESULT mad.gif
This only happen with some of the cDNAs tested, other cDNAs (I mean another samples) on the same plate work fine...

What does this high initial fluorescence mean?? I dont think its due to oversaturation right? 'cause once I used too concentrated cDNAs and only the Ct appeared earlier, the initial fluorescence was the same (or approximately) for every dilution.

Thanks for the help!! rolleyes.gif

-kukita-

first, is it repeatable for those samples?

second, are those particular wells in your machine contaminated with fluoresence?

-aimikins-

QUOTE (aimikins @ Oct 19 2007, 11:36 AM)
first, is it repeatable for those samples?

second, are those particular wells in your machine contaminated with fluoresence?


Yes, it is repeatable for those particular samples... and no, it doesnt appear to be any contaminated well in the machine...

-kukita-