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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