Digesting lambda prep

Robert Saunders 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.

Robert



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