cloning large fragments

P.J. Shaw pjs14 at le.ac.uk
Mon Mar 27 06:32:37 EST 2000


I routinely subclone inserts from lambda DASH clones. Usually, it's no
problem to get the whole insert cloned using the SalI sites in the
polylinker, if there's no internal sites. Don't bother gel-purifying; just
ligate an excess of cut fragments to bluescript which has been
dephosphorylated with SAP. Transform by electroporation and pick whites
(usually not a lot!). DH5a is OK, sometimes SURE cells are needed if you
get deletions.

Phil.

pjs14 at le.ac.uk  





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