Bacterial Plasmid DNA extraction

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lmenwe
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Bacterial Plasmid DNA extraction

Post by lmenwe » Sat Jun 16, 2007 7:59 pm

what are the usage of GTE buffer, potassium acetate, 70% ethanol, 95% ethanol and isopropanol in large scale extraction of plasmid DNA extraction

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canalon
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Post by canalon » Sat Jun 16, 2007 9:43 pm

GTE Buffer?

As for the other, they are used to precipitate and wash DNA. Usually if you use isopropanol, you do not need 95% ethanol since they both serve for the precipitation. 70% ethanol is just a washing solution that will not resolubilize DNA, but still rinse out the Acetate
Patrick

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lmenwe
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Post by lmenwe » Sun Jun 17, 2007 5:48 am

sorry it's TE buffer. I had made a mistake. Can you please explain the detail of them. I need to answer these in my report. If can please give the link for me thanks.

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Post by canalon » Tue Jun 19, 2007 6:50 pm

TE is TRis EDTA. Tris is a buffer, EDTA is an chelator for divalent cations. Since many DNAse use metallic cations (Mg2+ etc..) as coenzyme, if they are chelated, they will not be able to degrade your DNA during extraction.
Patrick

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