Dear [log in to unmask],

I have to correct my earlier statement in an previous email to the 
bulletin board (see below): One of the detection channels of the AB 
StepOne is in the right detection range and the signal is strong!

Our lab just purchased the new AB StepOne RT-PCR machine for conducting 
the thermofluor experiments.
I tested it before buying and it works just fine.

In Europe, AB has a trade-in running at the moment: If you can supply an 
old, even broken, simple PRC machine (not even RT-PCR), you get a 25% 
rebate which takes the price including tax below 19 k€. This could be 
interesting for anybody intending to buy a new machine.

- J. -



-------- Original Message --------
Subject: 	Re: Summary of Thermofluor
Date: 	Thu, 14 Feb 2008 12:05:08 +0100
From: 	Jeroen Mesters <[log in to unmask]>
To: 	[log in to unmask]
References: 	<[log in to unmask]>



mesters wrote:
> Sorry for the off-topic but can somebody recommend highly a sensitive
> RT-PCR machine for the thermofluor experiment (sypro orange).
> That would imply excitation below 500 nm (ideally 470) and detection
> at about 570 nm, right? I know several simple machines have a problem
> with the 570 nm...
>
> Jeroen.
>
Many thanks to all that commented to my query!

A list of RT-PCR machines that match the specifications (ex. 470 and
det. 570 nm (!), melting curve measurement possible, listed according to
price) are:
a) Biorad MiniOpticon
b) Stratagene Mx3005p
c) Biorad Biorad iQ5 and Aplied Biosystems StepOnePlus

Other RT-PCR machines (not exactly the right detection filter but
sufficient signal in the channel: listed according to price):
a) Aplied Biosystems StepOne
b) Biorad Biorad myiQ (rumors say it can be supplied with the correct
filters. If so, it is slightly cheaper than the Mx3005p but clearly less
flexible)

The Stratagene Mx3005p (beware, the Mx3000p is not useful at all) is the
most flexible machine for the money (5-channel detection among other
things) and has been used successfully used with Sypro orange. Also the
Biorad machines (iQ5 but also MiniOpticon) have a respectable reputation.

Looks like the Biorad MiniOpticon (2-channel machine) will fit my budget
but I must test the Applied Biosystems StepOne (3-channel machine, best
offer) before making the final decision. This has to do with the fact
that if the detection filters have a small band-path, a machine with an
off-peak but broad band-path filter may detect more signal at the end of
the day.

- J. -





-- 
Jeroen Raymundus Mesters, Ph.D.
Institut fuer Biochemie, Universitaet zu Luebeck
Zentrum fuer Medizinische Struktur und Zellbiologie
Ratzeburger Allee 160, D-23538 Luebeck
Tel: +49-451-5004070, Fax: +49-451-5004068
E-mail: [log in to unmask]
Http://www.biochem.uni-luebeck.de
Http://www.iobcr.org
Http://www.opticryst.org
--
If you can look into the seeds of time and say
which grain will grow and which will not - speak then to me  (Macbeth)
--