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?what is better for protein precipitation ammonium acetate or sodium chloride in

DNA extraction protein precipitation amonium acetate sodium chloride

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#1 JessGarcitocromooxidasa1



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Posted 09 July 2019 - 11:02 AM

Hi everyone
I'm student and i am comparanding two protocols of DNA extraction. One of them used ammonium acetate for protein precipitation and the other one sodium chloride. my question is what is better and why?

#2 OldCloner



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Posted 11 July 2019 - 07:26 AM

As I recall, sodium chloride itself can co-precipitate with the DNA in the last ethanol –ppt  step, leaving you with “salty” DNA pellets. So my mentor of long ago recommended either sodium or ammonium acetate, which are less likely to co-precipitate. However I must admit to using commercial kits for most of the last years of my career.  What kind of DNA are you extracting (plasmid, genomic?) and from what organism? ( Gram neg. or gram pos. bacteria, mammalian cells, tissues, plants?)  What’s the lysis protocol?  At what step are you adding the salt?  All these thing might matter to the answer.

#3 iosman987



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Posted 21 March 2020 - 05:49 AM

That won't work- because if you are adding equal amounts of four 100mM solutions together, you are increasing the total volume by a factor of 4 and therefore diluting each dNTP 1/4.

What I do is add 10ul of each dNTP then make up to 100ul with nuclease free water for a 1/10 dilution. Thanks

#4 bob1


    Thelymitra pulchella

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Posted 21 March 2020 - 01:17 PM

@iosman987 - I think you answered the wrong question...

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