In article <199404191228.AA06001 at ua.d.umn.edu>, "Dave Eide" <deide at ua.d.umn.edu> writes...
>Buddies,
>>We would like to do some one-step gene disruptions using flanking regions
>smaller than the recommended minimum. Rothstein (Meth. Enzymol. vol. 194)
>suggests using flanking regions of 100 bp or more on each side of the selectable
>marker and the method we want to try gives us only about 50 bp on each side.
>Does anyone have information regarding homologous recombination of fragments of
>this size? Any input will be greatly appreciated.
>>Dave Eide
>Dave,
Actually it works with even shorter homologous regions. Check a
protocol that uses PCR in NAR: 21(14), 3329-3330 (1993). This protocol have
been used in our lab with good results. (used just once).
Good luck,
Jaime E. Arenas
Division of Biology
CALTECH
