Basis of gene targeting efficiency

Josef P. Magyar magyar at
Thu Nov 26 04:54:57 EST 1998

I have been working with gene targeting for a while and have seen
constructs workin good (homologous recombination frequency (HRF) 1:5 to
1:20), less good (HRF=1:200 down to 1:5000) and such which never worked
though constructs were correct.
Now I believe to know, what the reason is for such differences.

I would like to receive from you constructs, which you have used for
gene targeting in order to compare them. It does not matter, whether
they worked good or bad (even those, which were bad are hard to find,
since they never come to publication). If you have concers due to
concurrency or so, send me please SauIIIa digested constructs containing
the plasmid (and only that) you used for gene targeting. For the
analysis, I would need about 10µg of your DNA in a tube (would be nice,
if I would not need to make DNA prep for each sample  ;-)   )

Please indicate the following along with your samples:

Your name and institute: as it should appear in acknowledgement
Your address and eMail: just in case there would be some further
Name, concentration (or amount) and HRF of your plasmid  DNA
If your DNA is digested with SauIIIa
Reference for the plasmide (if it exist)
If possible a map if not, at least the following (as an example)
      Linearised vector looked like:
            short arm (1kbp) / NeoR (1.2kbp) /long arm (4kbp) / HSVtk
(3kbp) / vector (3kbp)

In case of further questions, mail me to      magyar at

Thanks in advance
Josef P. Magyar

Postal address:
Josef P. Magyar, Ph.D.
Institute of Cell Biology
Swiss Federal Institute of Technology
ETH-Hoenggerberg, HPM F27
CH-8093 Zurich, Switzerland
Tel: +41 / 1 / 633-33-54
Fax: +41 / 1 / 633-10-69
eMail: magyar at

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