Growing Up TA Cloning Products

Duncan Clark Duncan at genesys.demon.co.uk
Tue Aug 30 05:27:46 EST 1994


In article: <33qe76$5rt at agate.berkeley.edu>  brunet at uclink2.berkeley.edu (Lisa J Brunet) writes:
> 
> Andrew McArthur (AMCARTHU at UVVM.UVIC.CA) wrote:
> : Hello everyone.  For those of you that use the TA cloning kit (Invitrogen),
> : what is your prefered method for overnight cultures of your cells with
> : inserts?  I have been using an overnight in LB (with ampicillin) but have
> : not been getting a satisfactory amount of plasmid DNA.  Has anyone tried
> : TB media or chloramphenicol with this kit to increase plasmid DNA yield.
> : Much thanks for your comments.
> 
> : Andrew
> Yeah, me too. You'll notice that the vector is based on pBR322 so it 
> grows like crap. You can amplify it or just get enough DNA so you can 
> subclone into something else. That's what I did.

I use TA all the time and don't have a problem with yields. It is pUC 
based with pUC copy numbers. Only if the gene is expressing and toxic 
do I have problems. Using alkaline lysis on 1.5ml O/N cultures in LB I get 
several ug plasmid. TB gives about double.

Duncan

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Duncan Clark                        | Internet:    duncan at genesys.demon.co.uk
G4ELJ                               | Compuserve:  100015.1406 at compuserve.com
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