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Perneseblue's Corner



E. coli ligase is not the way

Posted by perneseblue, 01 September 2010 - - - - - - · 14,668 views
multiway ligation
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 warn that...


Ligation 5 way and beyond

Posted by perneseblue, 27 August 2010 - - - - - - · 802 views

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 could...





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