Expand PCR and TA cloning kits?
jpcd0 at mole.bio.cam.ac.uk
Mon Apr 1 07:20:56 EST 1996
In article <4jeubs$r7c at news.duke.edu>, Margon Vandongen
<vando006 at mc.duke.edu> wrote:
> I have no experience with that particular enzyme but we use a mixture
> of PFU andd Taq polymerase which I think is exactly the same as these
> companies market for a much higher price. I posted this a little while
> back but here is a repeat: in our hands "mainly A's" meant no luck in TA.
> We now routinely spin the sample through a Qiagen column, change to a
> Taq buffer, add dntp's and taq and leave it at 72 degrees for ten
> minutes.It is more expensive this way but I prefer proofreading and it
> works every time!! Good luck,
Dont you have to purify the nukes away after this, prior to ligation?
Perhaps pfu is different to pwo as the other reply to my question said
Our group has used Expand Taq and successfully cloned into Invitrogen's
TA cloning vector on a regular basis. Cheers. Barry.
John Dixon Lab 44 (1223) 334131
Wellcome/CRC Institute Fax 44 (1223) 334134
Department of Genetics
United Kingdom e-m: jpcd0 at mole.bio.cam.ac.uk
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