RE: Quantitative PCR & Housekeeping Genes

From: Van Bockstaele, Dirk (Dirk.Van.Bockstaele@uza.uia.ac.be)
Date: Wed Jun 07 2000 - 10:55:49 EST


Hi Mario,

At the moment we are amplifying the abl message and found the information
present in "Debate Round-table: Multiplex PCR for quality control of
template RNA/cDNA in RT-PCR assays.  F Watzinger and T Lion, Leukemia year
1998, page 1984-1986 (can't find the volume reference)" very useful: these
people report on the use of bcr, abl, b2-MG and PBGD (porphobilinogen
deaminase) in a multiplex assay!

Since I'm also just recently engaged in this kind of work I would appreciate
receiving a copy of the responses you will get.

Best Regards,
Dirk


Prof. Dirk Van Bockstaele, PhD
Laboratory of Hematology
Head Flow Cytometry
Antwerp University Hospital
Wilrijkstraat 10
B-2650 Edegem
Belgium


> ----------
> Van:	Mario Roederer[SMTP:Roederer@drmr.com]
> Verzonden:	dinsdag 6 juni 2000 22:18
> Aan:	Cytometry Mailing List
> Onderwerp:	Quantitative PCR & Housekeeping Genes
>
>
> Hi, a little off the flow topic (but only a little)....
>
> we are doing quantitative PCR on very small numbers of sorted cells,
> and are looking around for a different housekeeping gene for control
> purposes than beta-actin.  We need one with an intron so that we can
> differentiate cDNA from genomic in our ultrasensitive amplifications.
>
> For people who are using housekeeping RNA's as controls, I'd like to
> get feedback:  what gene products do you measure?  Do you take
> advantage of spanning a splice junction?  Have you measured the
> variation in different cell types or stimulation conditions?
>
> Thanks for any info
>
> mr
>



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