transgene insertion and repetitive sequences
(by ucgatan from ucl.ac.uk)
Fri Aug 17 09:30:46 EST 2007
On Thu, 16 Aug 2007, Ed Siefker wrote:
> Tom Anderson wrote:
> > Once you've got that far, order up a genomic clone (or several) from
> > normal mice covering the area of the insert and use it as a probe on a
> > Southern blot. Comparing the pattern to that from normal mice will
> > tell you with, i think, a high degree of precision where your insert
> > is.
> I'm not sure I understand this. We do know that the transgene is in
> band 3A3. If I get a BAC from this region, how would I use it as a
> probe? It's kind of big for that isn't it?
Yes, i suppose you do really need to get to substantially sub-band
resolution to be able to do it this way - unless you have a very small
band or are willing to work with 50-100 BACs.
Does anyone sell per-band BAC collections in 96-well plates?
> > Another way of getting rid of real X DNA would be to find some
> > genomics people and get them to flow-sort chromosomes from your mice.
> > Once people have things set up for doing this, it works very well.
> Hey neat, I didn't know they could do that.
Nor did i until a friend started a PhD on chromosome biology, and did it!
Tom Anderson, MRC Laboratory for Molecular Cell Biology, UCL, London WC1E 6BT
(t) +44 (20) 76797264 (f) +44 (20) 76797805 (e) thomas.anderson from ucl.ac.uk
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