Liquid vs solid using Invitrogen TOPO kit

Frank O. Fackelmayer fof1 at chclu.chemie.uni-konstanz.de
Thu Jul 9 10:05:51 EST 1998


Hi Mark,
could you please provide us with a recipe for your TSB. Sounds promising
Frank

Mark Denhart wrote:
> 
> Hello,
> 
> I routinely use the kit from Invitrogen with great success.
> 
> However, for ease of preparation I use Tryptic Soy Broth instead of LB
> or SOC.  TSB is much easier to prepare and the cells are happier.  The
> colonies grow at least 5X larger on the selection plates (you only need
> to use 50 ul of cell solution per plate) and I get  4 to 6 ug of
> purified plasmid per mL of liquid culture (using a 10 mL per flask).  I
> use 50 ug/mL of Kanamicin Sulfate.  Both pCR2.1 and pCR II can provide
> both Kan and Amp resistence.
> 
> The cells that come with this kit are either INFalpha' or TOP10 cells.
> I have used both with equally good results.
> 
> Mark A. Denhart
> Research Analyst
> American Water Works
> Belleville, IL
> (618) 235-3600  x255



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