Elution

The proper way to make negative control is to set up a ligation of your digested vector with water (or TE buffer/elution buffer you are using) in place of insert DNA and use that ligation as negative control in transformation. That gives you an idea of the background (self-ligated ...

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... first time ı see a value like this. But ı make dna cleaning and concentrating prosedure and forget my dna sample 5 or 7 minutes in the final step(elution step). Is this the matter I really don't undestand??

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... vanishes : it's there in the soluble extract that i load on the column, but not anymore after the column, neither in the flow through or the elution... I've been then trying the old batch method... In parallel, with Talon resin (cobalt) and Ni-NTA resin (nickel). What was weird, was a change ...

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... an in-vivo method to try. I've noticed here: http://www.sciencedirect.com/science/article/pii/0014579383806243 The use of blue-sephadex, Ap5A elution and a second G-100 sephadex separation yield homogenous adenylate kinase in E.Coli anyway (the putative gene i'm dealing with show's a high ...

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Well from what I understand, 66mM of imidazole was enough to elute your protein so you do not need to add more, and you can use your elution to do all your experiments further. I would agree, but he wrote, that the first time, he had the protein also in franctions 80 and 100 mM

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Elution

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Cloning/Transformation question

Re: dna purity is 17? what does it mean?

Cobalt turning orange ??

Re: T7 Expression System

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Elution

Results from our forum

Cloning/Transformation question

Re: dna purity is 17? what does it mean?

Cobalt turning orange ??

Re: T7 Expression System

Re: