Adding 3' terminal thymidine to blunt ended DNA

Chi-kuang Wen ckwen at
Fri Jul 5 12:39:36 EST 1996

Simon Q.J. RICE ("SimonQRICE%notes" wrote:
: -Vectors which improve the efficiency of subcloning PCR fragments are 
: fairly common place these days.  They are made by adding a 3' terminal 
: thymidine to a blunt ended vector fragment taking advantage of the 
: non-template dependent addition of a single deoxyadenosine to the 3-end 
: of PCR products

: Anybody any idea how the thymidine is added to the 3'-end ensuring only 
: one thymidine is added at a time?

Just add dTTP, your blunt-ended vector, Taq, buffer at 72 hr to let the Taq to
do its job.


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