Large plasmid cloning
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Tue May 4 18:24:54 EST 2004
In article <4097e869$1_1 at mk-nntp-2.news.uk.tiscali.com>, "N. Horner" <horner1978 at yahoo.co.uk> wrote:
>I've been attempting some cloning since January with no luck. It turns out
>that the Ligase I was using was inactive (Promega). You might want to check
>yours by ligating Lambda DNA/Hind III fragments and then checking on a gel.
The fact that one has to use everything "Promega" at twice higher
concentrations aside, the most common problem with it's ligase
is bad/hydrolysed ATP in ligase buffer. Adding extra 2 mM ATP
usually solves the problem.
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