5 Minute Ligation
polli~j at glaxo.com
Mon Jan 9 17:17:18 EST 1995
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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?
>University of Manitoba
> 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.
>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 ??)
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!
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