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perneseblue's Photo

perneseblue

Member Since 26 Jan 2009
Offline Last Active Jul 09 2012 04:45 PM

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Community Stats

Group Global Moderators
Active Posts 530
Profile Views 18,279
Member Title Unlimited ligation works!
Age Age Unknown
Birthday Birthday Unknown
Gender
Not Telling
Location
USA (formerly UK)

About me

My research interests
synthetic biology

Contact Information

Website URL http://

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Blog Entries

Perneseblue's Corner > E. coli ligase is not the way

Posted 01 September 2010

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...

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Perneseblue's Corner > Ligation 5 way and beyond

Posted 27 August 2010

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...