IUBio

Fluorescense quenching

Denis Bergeron denisb at algene.com
Sat Mar 20 20:20:05 EST 1999


We have observed that the best fluorescence intensity of your product are
beetween 200 and 3000 (These is the machines best parameter). Somehow when
the product gets too intense, the peaks looses resolution and becomes
"fatter", so you are loosing precision on your data. Be careful when you
accept these results, make sure that your fragments are well sharp and
define.
You are right by saying that about 4000 it starts to degenerate but keep
in mind thar the genescan software might give you a 4000 peak height when
the real peak height might be 6000. The software in this case, compensate
by showning you a larger peak instead than a sharp define peak.



In article <7cmqkk$sdo at net.bio.net>, "Mark E. Berres"
<markb at ravel.zoology.wisc.edu> wrote:

>Does anyone have any knowledge concerning the maximum
>amount of fluorescense an ABI 377 can detect and still
>maintain a linear relationship between the mass of
>fluorophore labelled DNA and fluorescense intensity?
>
>In my experience, it seems to be somewhere around 4100
>(rfu) but this is only empirical.
>
>                        Thanks!
>
>                                Mark




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