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E. coli ligase is not the way

Dear Blog,

It has been a case of "I did warn you."

I have attempted to improve the efficiency of my multiway ligation by using a different ligase, a higher fidelity ligase. The only other ligase sold commercially is E.coli ligase. It is stated that E. coli does have higher fidelity compared to T4 ligase, however NEB did...



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Ligation 5 way and beyond

Dear Blog,

I have decided to find the limits of multi-way ligation. Thus far, I have determined that the techniques I am using gives me a reproducible limit of 6-ways. 7-way ligation occasionally works. And all attempts at 8-way ligation have ended in failure. My current screen size is 96 colonies..

At this point there are two obvious ways I...



February 2012

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