Real time PCR. Funky fluorescence?

[Gilbride, Kevin]

I saw this once before while using ABI7700 and it turned out to be
"falling Rox". The reference dye for some unknown reason loses its
fluorescence and without the reference the target signal plummets.
JMHO
Good luck,
K 

-----Original Message-----
From: methods-bounces from oat.bio.indiana.edu
[mailto:methods-bounces from oat.bio.indiana.edu] On Behalf Of Matti Savoila
Sent: Wednesday, April 23, 2008 7:47 AM
To: methods from magpie.bio.indiana.edu
Subject: Real time PCR. Funky fluorescence?

Hello everyone!

I am doing some real time PCR with the LightCycler 1.5 instrument, but
I'm getting weird results. The fluorescence rises, like it should, but
suddenly in one cycle it plummets down. Examples:
http://www.mv.helsinki.fi/savoila/d_kuvat/Nayte_4_edit.jpg
http://www.mv.helsinki.fi/savoila/d_kuvat/Nayte_6_edit.jpg

Have you seen behaviour like this, and more importantly, have you seen
a solution to problems like this? Number 10 in both samples has been
amplified normally, which is strange. If somebody has some ideas, I'd
be thankful.

Best regards
Matti Savoila

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