Large Scale Ligation.

Patrick F.H. Lai pfhlai at LOOKSMART.COM
Mon Aug 10 20:12:52 EST 1998


---- Begin Original Message ----
>Hello all
>    I have a question that I can not find any reference to,I am >hopin=
g that
>some one in this group can give me some insight. I need to do a >ligat=
ion,
>where I need mg of the ligated product. There is potential problems >a=
s I see
>it in the amount of ligase needed for one things, is there any >refere=
nce out
>there regarding parameters of doing such large amount of ligation ?
>
>-Thanks in advance
>T.L
>--
>Tam Anh Le
>Clinical Neuroscience
>St Vincent Hospital Melbourne.
>
---- End Original Message ----


Dear Dr. Tam Anh Le,

How about inserting your ligation product into a vector first ?
You only need a small scale ligation to start with.
You can then release as much as you need by restriction digestion of
MaxiPreps, and then gel extract your fragment.
Once your ligation product is in a vector, you can even easily do
sequencing to check if your ligation was alright.


Good Luck !





Hope this is useful info.   :-)


Patrick F.H. Lai  < PFHLai at looksmart.com >
Graduate Student
University of Toronto
Toronto, Ontario, Canada





       
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