Large plasmid inserts, electroporation

Stephen Smith sgsmith at tcd.ie
Thu May 4 15:15:34 EST 2000


Hi,
As Warren Gallin pointed out it would be better to electroporate large
plasmids and he's quite right.

In the following paper the authors studied the relationship between plasmid
size and electroporation efficiency, they got efficient electroporation
even with a plasmid 136kb in size!!

Leonardo ED & Sedivy JM
A new vector for cloning large eukaryotic DNA segments in Escherichia coli.
Biotechnology (N Y) 1990 Sep;8(9):841-4

However, its probably better to ligate into a low copy number vector. In
the past I could successfully clone 25kb in a low copy number vector
(pLG339 copy number 4), but I could'nt clone this fragment in bluescript.

Regards

Stephen


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