RNA/DNA Co-extraction. Which Phenol pH?

Use this category for the exchange of ideas, methodologies and references regarding the isolation, manipulation and analysis of RNA. (Extraction protocols, Northern Blot analysis, RNase Protection, Differential Display, In Vitro Transcription, etc.)

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RNA/DNA Co-extraction. Which Phenol pH?

Postby nouse » Mar 07 2017 7:00 am

Hi,

we are planning a parallel sequencing of rRNA and rDNA.

We are using a PCI-based extraction. It is my understanding that PCI pH 8.0 let both NAs migrate to the aqueous phase, whereas acidic PCI precipitates DNA, leaving RNA. Is that correct?

Which approach would you then recommend:

i) Co-extract with PCI at pH 8.0, aliquot, degrade DNA with DNAse.

or

ii) aliquot after lysis, and extract RNA with PCI pH 4.7 and extract DNA with PCI pH 8.0.

?
nouse
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