Hi Mario,
This might be even more off-topic, but...
Mark Wills (mailto:mrw1004@mole.bio.cam.ac.uk) in my department looks
for specific V beta rearrangements in his CTL clones and ex vivo
blood samples, and has the advantage of being able to compare that
to TCR C region, ratioing the two gives him an idea of clone size.
He wrote the method up in J Immunol 1999 Jun 15;162(12):7080-7
"Human Virus-Specific CD8+ CTL Clones Revert from CD45ROhi to
CD45RAhigh In Vivo:CD45RAhiCD8+ Tcells Comprise Both Naive and Memory
Cells".
Maybe you could use a similar strategy if there's a constant splice
junction on your cDNA that you could compare with a variant of
interest (I presume that the technique would apply generally to Ig
superfamily members and CD45 to some extent),
Ray
>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
--
Ray Hicks
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