Re: Misc.:"Taqman"

Ross Durland (durlar@genemedicine.com)
Tue, 2 Feb 1999 11:21:34 -0600

We have several years experience running TaqMan qPCR assays on the 7700.
Most of our work is for DNA, but we've done some qRT-PCR as well. In my
opinion, if you have lots of samples, need high throughput & reliable
quantitation, & can afford it, the 7700 is the only choice. In our hands,
for a well-behaved qPCR assay, intra-assay precision of calculated
quantities can be as low as 5-10% CV. Run-to-run reproducibility for a
reference control sample can be as good as ~ 11% CV. These values do not
include variability from DNA (or RNA) isolation, or from reverse
transcription, which is often substantial.

Drawbacks of the 7700 are that it's big & heavy & requires 220V, so you
can't put it just anywhere. Also, instrument reliability has not always
been the best. I recommend a service contract after the warranty expires.
Also, it only runs off PowerMacs. There are other occasional difficulties
as well, but the advantages far outweigh the disadvantages for us.

I don't know what cytokines & interleukins you want to monitor, but PE is
coming out with pre-developed qRT-PCR assays for a number of human & rodent
genes.

The best way to get lots of info is to join the 7700TAQMAN discussion
group. Send an email to listproc@listserv.acns.nwu.edu. The message body
should be the single line, "subscribe 7700TAQMAN." Post your questions
there, & you'll reach a good number of heavy users who can help you,
particularly regarding qRT-PCR. You may even find people who have developed
assays for your genes of interest.

Good luck!

Ross Durland
Director of Core Technologies
GeneMedicine, inc.
8301 New Trails Drive
The Woodlands, TX 77381-4248
281-364-1150
durlar@genemedicine.com

Scottie Adams <sadams@northnet.org> on 02/02/99 06:59:27 AM

To: Recipients of ABRF List <abrf@aecom.yu.edu>
cc: (bcc: Ross Durland/GeneMedicine)
Subject: Misc.:"Taqman"

Greetings
Does anyone have any experience with the Applied Biosystems
"Taqman" PCR 7700 system?? We are considering using this system to monitor
cytokine/interleukin production instead of the rather tedious RNase
Protection Assay or other quantitative PCR systems.
I would appreciate hearing from anyone on the subject.
Scottie
Happy Groundhog Day to all - especially Jim Bloom. Are gophers related to
groundhogs????
Scottie Adams aka Pamela Scott Adams
Molecular Biology Core Facility
Trudeau Institute
100 Algonquin
Saranac Lake, NY 12983
Phone: 518-891-3080
Fax: 518-891-5126
Email: sadams@northnet.org
http://www.trudeauinstitute.org