how CIP treatment is done - (Jan/17/2010 )

Dear all ,
I am just wondering how CIP treatment is done since I've never tried it before .Do we always need to purify the product after CIP treatment , what if the concentration of vector is too low .


Regards
Sandeep

sahil on Jan 17 2010, 03:43 AM said:

Add some CIAP to your reaction with the appropriate buffer, incubate for a while at 37 deg C.

Yes, otherwise it will remove phosphates from your other DNA (insert or plasmid) when you mix the two.

Make some more - you have glycerol stocks right?

bob1 on Jan 18 2010, 08:24 AM said:

the same my situation. The concentration of vector ( M13) is very very low, it was un-visualized on agarose gel (0.8%) but there are a few because i try to transformant and get a few colonies. How can i resolve these problems. Many thanks for any suggestion

you could try Antarctic Phosphatase (http://www.neb.com/nebecomm/products/productM0289.asp) which can be heat inactivated removing the need for purification- however I think you should go with bob's suggestion and just make some more- cheaper, and faster, to do it that way