TA Cloning Vectors - Reply

Sun Feb 19 04:01:38 EST 1995

>Hi Klas,
>a related question,
>do you know what would be the optimal phasing in order to create a TA cloning 
vector such that the resulting
>cloned PCR product would be (frequently) in a particular reading frame?
>ie can one assume that a single T predominates on the vector and a single A on 
the PCR product?
>Cal Vary

Hi Cal and netters

We have been sequencing a couple of ligations with two T:s but in most cases a 
single T is found. So, yes, you can assume that a single T predominates but 
using it for in-frame ligations for expression will involve sequencing 
your ligation site. However, if you want to be sure of having one T on a vector 
I think that most commercials vector use an enzyme which will create a single 

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 I take a deep breath and get real high
 and I scream from the top of my lungs
 Whats going on?"


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