5 Minute Ligation

[Joseph Polli]

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In article <84652160105991.440720 at usa>,
        polli~j at glaxo.com ("Joseph Polli") writes:
>[This message is converted from WPS-PLUS to ASCII]
>
>Dave Boyd wrote:
>Anybody try Boehringer Mannheim's 5 minute ligation kit yet?
>Whats the secret ingredient? Does it really work?
>
>Dave Boyd
>University of Manitoba
>
>
>Dave,
>
>    I have used the Boehringer Mannheim's 5 min. ligation
>for several months now and it has worked very well.  I
>think you should give it a try.
>
>Joe Polli
>Dept. Molecular Biology
>Glaxo Research Institute
>RTP, NC  27709


>what kind of efficiencies do you get ?   do you have to use 
>bigger amounts of DNA ??   would it be suited for any kind of 
>ligation (like when making a library for example ??)

>thanks                  paolo  


Paolo,

    I have not done the experiments to answer your question on 
efficiency.  I did not modify the amount of DNA I had been using 
for previous ligations (40 ng vector/ 25 ng insert generally).  
So far we have only used the 5 min ligation kit for routine 
subclonings.  Folks in the lab  were interested to see if it 
worked well, and so far it has worked well.  I can't comment on 
its use for making libraries, but as Dave Micklem pointed out in 
a previous message that libraries are alot of work and for now I 
would spend the extra time with what has previously worked.  What 
I would like to see developed is the bacterial strain that will 
grow so that your colonies are ready in 5 minutes!

Joe