Expand PCR and TA cloning kits?

John Dixon 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:

> John,
> 
> 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, 
> 
> Margon 

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

John,
 Our group has used Expand Taq and successfully cloned into Invitrogen's 
TA cloning vector on a regular basis. Cheers. Barry.

Thanks anyway.

-- 
John Dixon                                        Lab 44 (1223) 334131
Wellcome/CRC Institute                            Fax 44 (1223) 334134
Department of Genetics
Cambridge University    
United Kingdom                           e-m: jpcd0 at mole.bio.cam.ac.uk



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