cloning large fragments

Chris Boyd cboyd at holyrood.ed.ac.uk
Mon Mar 27 06:41:46 EST 2000


Jean-Christophe Billeter <9902301b at student.gla.ac.uk> wrote:
: Does anyone have expertise in sub-cloning big fragments (i.e.12-15kb) from
: BACs into
: conventional high-copy number plasmids e.g.bluescript? What kind of band
: purification
: method? What insert : vector ratios? additions/improvements in ligations?
: What kind of
: E.coli strain would you use for transformation? Furthermore, I would be
: interested in
: getting feedback about the biggest insert someone has managed to sub-clone
: into a
: plasmid.

See

   Tao, Q. and Zhang, H. B. (1998) `Cloning and stable maintenance
   of DNA fragments over 300 kb in Escherichia coli with conventional
   plasmid-based vectors.' Nucleic Acids Res., 26, 4901-4909.

and weep.


Best wishes,
-- 
Chris Boyd                      | from, but \    Medical Genetics Section
Chris.Boyd at ed.ac.uk             | not for,  /    MMC, Edinburgh Uni.,
http://www.ed.ac.uk/~cboyd                       EH4 2XU, SCOTLAND




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