YAC sub-cloning

Ola Myklebost olam at radium.uio.no
Thu Apr 15 11:02:46 EST 1993


In article <1993Apr6.095011.17350 at gserv1.dl.ac.uk>, acurtis at crc.ac.uk (Mr.
A.R.J. Curtis) wrote:
> 
> 
> We have been attempting for several months to subclone a 450kb YAC to
> make representative libraries:
> a) SauIIIa cut small-fragments cloned in a plasmid vector
> b) SauIIIa partial digest cloned in a cosmid vector

> In our hands the starting material is so sparse or gets degraded.
> Our YAC starts as a band cut out of a LMP-agarose pulse field gel.


I have no personal experience, but the problems you describe are typical.
at the Cold Spring Harbor course on YAC analysis (which is highly
recommended), we were advised to rather make a total lambda library of
yeast with YAC and then screen for YAC clones with total human DNA and
vector ends. Such a library is easy to make and doesn't need to be large (I
believe a few thousand clones is enough).

Ola Myklebost             Email olam at radium.uio.no
Dept of Tumor Biology
Inst for Cancer Research
The Norwegian Radium Hospital
N-0310 OSLO, Norway
Tel +47-22-50-60-50, xtn 9830, Fax +47-22-52-24-21



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