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Insert direction in TA cloning


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#1 sara.r

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Posted 11 January 2015 - 12:01 AM

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



#2 bob1

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Posted 11 January 2015 - 01:33 AM

They can insert in either direction.



#3 phage434

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Posted 11 January 2015 - 07:31 AM

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.






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