RAG locus in mice

Josiah Wedgwood wedgwood at msvax.mssm.edu
Sat Oct 12 01:18:13 EST 1996

You have all the information you need.  The location of the introns is
usually conserved and for your purposes the exact sequence is
irrelevant. I would encourage you to use conserved sequences across
several species on two different exons.  This avoids problems of
mistakes in sequences in the database or use of a different strain of
mouse.  If you want to prove that your primers span and intron, then run
the PCR on genomic DNA, but use a long extension time because the
product you produce may be very large.

Josiah Wedgwood
Mount Sinai School of Medicine

Justin Reese wrote:
> Hello,
> I am currently a grad student at the University of Georgia, and am
> planning on doing some RT-PCR experiments for RAG expression.  I'd like
> to use primers which span an intron, so that I can differentate RAG mRNA
> signals from signals generated from contaminant genomic DNA.  The problem
> is, I cannot for the life of me find the genomic sequences for RAG1 and
> RAG2 in mice.  I have found genomic sequences for over 10 species, but
> not mice.  I'm beginning to think
> that this sequence has not been done in mice yet.  Does anyone know?  I
> have gotten the cDNA sequences for these genes in mice, and I've read 6-7
> papers about the RAG locus, but I can't seem to locate this information.
> I've also done computer searches for just about every permutation of RAG
> (and recombinase/ation activating/associated gene, etc etc).
> If anyone has this info, or knows of a way of obtaining it, please email
> me (jreese at cellmate.cb.uga.edu)
> Thanks in advance,
> Justin Reese
> University of Georgia
> Department of Cellular Biology
> jreese at cellmate.cb.uga.edu

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