A260 or Fluoromete ??? References

WSchick at aol.com WSchick at aol.com
Thu Mar 21 01:22:23 EST 1996


In a message dated 96-03-20 12:48:05 EST, bturner at med-itvax1.bu.edu (Bradley
Turner) writes:

>Would it be possible to post the references or methods for the in solution
>fluorescence assays of DNA and RNA that you mention below.  They sound
>interesting, and I might as well put my fluorimeter to use.
>
>For Hoechst, an application note with references is #119 available from
Hoefer/Pharmacia ( who also sell the 260 280 GeneQuant)  Both the single
wavelength DynaQuant (Hoefer's fluorometer) and the GeneQuant sell for about
$2500 in the US.
If you have current protocols in molecular biology vol 2, look up Gallagher,
S "Quantitation of DNA and RNA with absorption and fluorescence spectroscopy.
 Unit A.3.9.

For PicoGreen and OliGreen and the equivaletn SYBR greengel dyes, contact
Molecular Probes.  They even have SYBR DNA standards for gels.  Their FAX in
Eugene, OR is 541-344-6504  email is tech at probes.mhs.comuserve.com  I don't
see any references in their lit, but by now they should be appearing.  Their
Nov 1995 BioProbes 22 discusses these new dyes.   Note that the wavelengths
are different from Hoechst, so you could use your filter or grating
fluorometer.

An alternative to Hoechst is thiazole orange, and an application note
authored by Dan Simpson of Promega is available from Turner Designs  fax 408
749 0998, email mfeld at ix.netcom.com (Pam Mayerfeld).  For researchers wanting
a new fluorometer, they offer a quick change filter fluorometer (also offered
by Molecular Probes) for about $5000 in the US.  Advantage is that you can
change dye applications where you can't with the Hoefer unit.  Protease
assays, histamine, CyQuant cell proliferation, fluorescein, rhodamine, texas
red, etc.

Commercial attachment--I once worked at both Turner Designs and Hoefer, and
thus am biased toward fluorescence for many assays.  And I am still curious
about the comparison of 260/280/fluorescence for DNA.

Walt Schick



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