does anyone use Northern here?

Tim Spahlinger txs at po.cwru.edu
Fri Jan 11 14:41:03 EST 2002


Stefi,

Have you considered doing real-time PCR, ala TaqMan or Light
Cycler, to relatively quantify your 2 messages?  If you know
the sequences that you're investigating you might be able to
design 2 FRET probes whose fluorescence would indicate the
relative amounts of message.  The Applied Biosystems web
site (for TaqMan chemistry; www.appliedbiosystems.com) and
the web site for a company called Molecular Probes 
(www.molecularprobes.com; they have a handbook there that
you could look at) could help you determine if this is right
for you.  I'm not affiliated with either company but I am
familiar with their technology and reagents.

Stefi~ wrote:
> 
> Hi,
> I'm on the point of choosing Northern for my RNA analisys (read my last
> messages).
> Well, I have to demonstrate an accumulation of mRNA and the decay of the
> levels of a very similar mRNA (the proteins codificated by these two mRNAs
> are two GSTp). I have to design two probes that should discriminate between
> the two forms. What is the best lenght for a DNA probe marked with
> fluorescein?
> How many molecules of fluorescein is it better to insert in my probes to
> have a good signal for rare mRNA?
> Does anyone know how may I obtain commercially probes with more than one
> fluorescein per probe?
> Thanx a lot to averybody in advance.
> Stefano
> gifalone at tin.it
> http://web.tiscalinet.it/gifalone




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