3-Fragments direct ligation at one time??

Xinkun Wang xinkunw at hotmail.com
Tue May 1 16:16:40 EST 2001


Hi, there,

I am trying to ligate my promoter and another encoding gene into a different 
vector. So there are 3 parts to be ligated here. I had thought to ligate my 
promoter into the vector first, then ligate my gene into the half-finished 
vector. But recently I am told I can ligate them together at one time. It 
saves time and work. So I am planning to try it. In my plan, I want to cut 
my vector with HindIII and BamHI, cut the promoter with HindIII and EcoRI, 
and the insertion gene with EcoRI and BamHI. After putting them together, I 
hope they can join each other to make the vector with my desired two inserts 
in the right orientation. Since I never did it before, I am still concerned 
about the effeciency of this approach. Does anybody use this method before? 
If it really works, that would be great. Any suggestions are welcome.

Thanks in advance.

Xinkun Wang

______________________________
Xinkun Wang
125 Noble Research Center
Oklahoma State University
Stillwater, OK 74078
Tel: (405)744-6599
E-mail: xinkunw at hotmail.com

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