TA Cloning and screening

Darren M. Stauth stauth at crunch.usgmrl.ksu.edu
Mon Feb 27 10:23:05 EST 1995

In article <3iluaf$hk at mark.ucdavis.edu>, ez022056 at dale.ucdavis.edu (Edward
Wang) wrote:

> I am currently working on a TA cloning project.  We've looked through a 
> few "TA cloning kits" and noticed that Promega sell their kit at $66.00 
> and Invitrogen sells at appx. $200.00 for the same amount of kit.  
> According to the maps provided, the only two major differences between 
> the kits are 1. the multiple cloning sites and 2. no Kan res. in Promega 
> plasmid.  We have decided to purchase the Promega kit, but I would like 
> to know from people who have worked with either one or both kits if there 
> are major differences in protocol or efficiencies, etc.  
> Also, we plan to screen for inserts with Screen Test Recomb. Screen Kit 
> by Stratagene.  Has anyone used this kit and perhaps could make some 
> input as far as how efficient this kit is?
> All inputs are appreciated.

The Invitrogen is definitely the better kit in my experience.  It also
cantains the competent cells.  However to save money we are trying to apply
some of the Invitrogen procedures to the pGem-T kit such as (1) adding
B-mercaptoethanol to the comp. cells (2) Adding SOC instead of LB to the
transformation  (3) using fresh pcr products.  Hope this helps.

Darren M. Stauth  stauth at crunch.usgmrl.ksu.edu

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