Methods Digest, Vol 47, Issue 16

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Thu Apr 23 11:39:18 EST 2009


Dear Manish, 

As the DNA is not in the chloroform phase, it is in the aqueous phase which contains the phenol (i.e. the phenolic phase), too. Maybe I haven't expressed that clearly enough (I wrote "phenol phase" which of course is not correct, there is no separate phenol phase, of course.

Best regards,

Wo

-------- Original-Nachricht --------
> Datum: Thu, 23 Apr 2009 13:07:55 +0530
> Von: manish mishra <manishjnu8 from gmail.com>
> An: methods from oat.bio.indiana.edu
> Betreff: Re: Methods Digest, Vol 47, Issue 16

> Dear Wo,
> 
> As you have written
> 
>   "You might
> > remember: the DNA (and other water soluble stuff) is in the phenol
> > phase, lipids are in the chloroform phase and proteins are inbetween."
> 
> I am slightly confused. Is it really that DNA is in the phenolic
> phage? I think when we isolate genomic DNA by  Phenol: Chloroform:
> Isoamyl alchohol , the pH is around 8.
> 
> I think that nucleic acid should be present in aqueous phage at pH
> 8.The lower organic layer contains proteins dissolved by phenol and
> lipid dissolved by chloroform.
> I dont know at pH 8 whether RNA is in aqueous phage or at interface?
> 
> At lower pH the RNA is in aquous phage and DNA is in phenolic phage.
> 
> Am I correct. If possible kindly clear my confusion.
> 
> with regards
> Manish
> 

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