In article <7dvnb1$nrp at net.bio.net>,
Elizabeth Smith <ems217 at psu.edu> wrote:
> Hello,
> I am doing DNA sequencingn on a chip using an in-house built system with
> an Ar ion laser. I am currently using the ABI BigDye Primer sequencing
> kit and am not seeing any fluorescence from these dyes. ABI tells me the
> excitation wavelengths for these dyes are proprietary, so I don't know
> if I'm using the correct wavelength! I was wondering if anyone has used
> this ABI kit with an Ar ion laser or if you could suggest a better kit.
> Thanks,
> Liz Smith
>>
We are doing a SAGE project on human keratinocytes. We use the ABI-310 for
sequencing clones. As I understand it the Big Dyes from applied biosystems
(Perkin Elmer) all consist of a fluorescein donor and a dRhodamine acceptor.
So the excitation is , for all of them , a property of the fluorescein
acceptor : both the 488 and 514 nm lines of the (10mWatt) argon ion laser are
used. Emission A=dR6G=520nm , C=dRox=625nm , G=dR110=545nm , T=dTAMRA=595nm
In about a month or two we hope to get started with DNA chips too. Good luck
Gys de Jongh
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