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Yes, I have made it myself several times using pTZ57R, pGEMT and pBlueScript.

Please see this paper: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC333800/


In the paper, authors state that If you give equimolar amounts of dNTPs to Taq DNA
polymerase, dATP will be preferably added. However, if you supply only dTTP, then Taq
will add an extra T to the end of any blunt ended DNA fragment. Based on that you can
prepare your own T-vector using any vector backbone of choice.
Here follow a detailed protocol (as I usually do):
-1X Taq Buffer - final
-MgCl2 to 2.5 mM final concentration
-10 units of polimerase (yes, it is a lot)
-Blunt ended Vector (up to 5 micrograms)
-ultra pure water to 100 microliters.
Incubate for 1 to 2 hours at 72°C (test with your avaialble Taq)
Purify by silica column kit (or simply by precipitation) and quantify by nanodrop or any other
method.
Easy and cheap!
Other option:
If you are amplifying a target gene using polymerases that leave blunt ends, you can easily
improve your blunt end ligation by adding a few units of blunt end enzyme to your ligation
system. In my hands, EcoRV and SmaI cuts very well in the 1X Thermo Fischer ligase
buffer.
Link for the original idea: http://www.academicjournals.org/journal/AJB/article-full-text-pdf/
7EEAD3525311
P.S: Ensure that there are no cut site for the blunt restriction enzyme of your choice in the
target fragment to be cloned.
I hope things are clearer now.
All the best

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