Isolation of plasmid DNA - chemicals used-why?? (Nov/22/2006 )

could anyone help me out and explain why we use certain chemicals, some i know but am struggling with why during a miniprep wizard we use resuspension solution (Tris, EDTA, RNase), neutralisation solution (K acatate), and DNA purification resin?

Thanks.

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The resuspension solution is used to break the bacteria and to degrade RNA; the neutralization solution because when you add NaOH the pH increases and you need to lower it to 8 so that your plasmid DNA renatures and goes in solution while the genomic and the RNA can precipitate; the DNA purification resin is used, as the name says, to purify DNA

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Brilliant thanks, could you just confirm then that im right in saying TE buffer is used to protect the DNA from degredation??

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oh and poss a silly question but the column wash???? is that as simple as aying its just to was out the column/membrane in miniprep?

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Yes, EDTA chelates the bivalent ions that most enzymes need to work

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Yes, it is to wash the column

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