CIP or BAP
landel at helios
Fri Sep 16 09:28:18 EST 1994
In article <35bo43$2k5 at aragorn.unibe.ch>, <beblum at ubeclu.unibe.ch> wrote:
>RNA editing research group in Bern is
>requesting information about the use of CIP (calf intestinal alk. phosphatase)
>or BAP (bacterial alkaline phosphatase) in order to remove
>the 3' terminal phosphate from an RNA oligo. Which of the two works better for
>this special purpose?
>All information concerning the topic is appreciated.
The enzyme of choice CIP, because it can be heat inactivated by incubation
at 65 degrees, while BAP requires a phenol and chloroform extraction for
inactivation. CIP is *very* efficient in KGB (or "universal") buffer--simply
add a few units at the end of your digestion and incubate for 5 more minutes
at 37 degrees, then heat inactivate. Be careful not to incubat much longer,
since CIP does have a problem that it seems to chew up the ends of your
DNA if you let it go too long, making your fragment unclonable.
More information about the Methods