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Yu Zhang's Content

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#178476 Plasmid purification for Gibson Assembly

Posted by Yu Zhang on 09 August 2016 - 09:22 AM in Molecular Cloning

Dear all,

I am Yu, working in biology and biotechnology.

Now I have already obtained plasmid with an amount of 4 ug. What I gonna do is to double digest this 26 kb plasmid with XbaI and BamHI. Finally I will get two fragments, one 22 kb that I will take for my Gibson Assembly vector, and trash the other 4 kb. But I am facing with a problem. I have no idea about plasmid purification, that is to say I do not know how to separate enough amount of purified 22 kb vector. If run on agarose gel, I am not sure the High Pure PCR Purification Kit (Roche) will work well on this frament with so big size. Does anyone have good and detailed protocol on big plasmid purification? Can share your instruction will be so helpful.

Thanks in advance!





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