Digesting lambda prep
r.d.c.saunders at dundee.ac.uk
Thu May 22 11:06:22 EST 1997
Mart Speek wrote:
> : Mario Pineda wrote:
> : > I have done various phage preps in an EMBL3A Library and have yet to be
> : > able to cut my insert out of the phage.
> : In my experience, the golden rule is to always use liquid lysates!
> : There are many possible problems with DNA extracted from plate lysates.
> : Robert
> $$$ Hello Robert,
> How're you doin'? (sorry it is not a chatboard!)
> JUST to add to the Mario's question: lambda liquid lysates are generally
> difficult to make, i.e. require careful adjustment of phage to host ratio
> to maximize the yield. I would recommend PCR amplification of an insert
> provided flanking primers for EMBL3 vector are availabile. Good luck!
> Mart (in Estonia)
Mart (Hi, I'll email you!)
I dispute that liquid lysates are difficult to do. Just take a bit of
care over the multiplicity of infection, and away you go. Plenty of
information around on that! Saves a lot of time over the multiple
attempts to get good DNA from plate lysates. I wouldn't go for
amplifying an entire EMBL3 insert, personally.
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