Insert direction in TA cloning - (Jan/11/2015 )

HI 

I am a little confused about the TA cloning. when we clone an insert in TA cloning vectors which have an overhang T on their 5 site, are all the inserts in 5 to 3 direction or still there is the possibility that the insert orientation be in reverse like what we have in single enzyme cloning? 

Cheers

They can insert in either direction.

The reason is that both ends of your dsDNA fragment have a 5' end, and a 5' A is added to both ends. You just tend not to think so much about the other strand of the dsDNA, and are biased in your thinking in a way in which the chemical reaction is not.