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Tips for genomic DNA extraction from gram -ve bacteria


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#1 Nrelo

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Posted 23 February 2009 - 11:02 PM

I am new to genomic DNA extraction and seeking advises. Are there any tips? I am rather unsure about removing protein by phenol:chloroform, how much volume (e.g v/v ratio?) should I add into the lysate?

#2 hanming86

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Posted 24 February 2009 - 06:28 AM

View PostNrelo, on Feb 23 2009, 11:02 PM, said:

I am new to genomic DNA extraction and seeking advises. Are there any tips? I am rather unsure about removing protein by phenol:chloroform, how much volume (e.g v/v ratio?) should I add into the lysate?


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#3 AquaPlasmid

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Posted 24 February 2009 - 08:03 PM

You really don't need to use phenol/chloroform these days. Gram positive bacteria are tough to lyse, but with a combination of lysozyme, beadbeater, and sonicator to break up the cell walls in AquaGenomic or AquaRNA solution you will get the DNA you need.




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