Assuming I would like to use some chemical mediated process of precipitating RNA and polysacharides (from tissue lysate) but leave genomic DNA intact in the aqueous layer. What are the best methods to it?
NOTE: The main aim is to get rid of RNA contamination during genomic DNA isolation WITH OUT USING ANY RNASE/ENZYME BASED methods.
Hi
What is your downstream application? Will it actually be affected by the presence of RNA?
If you really think RNA to be interfering with your reactions, you can re-purify your extracts with alkaline-Phenol-chloroform method. Alternatively you simply run your extracts on an 0.8% agarose gel, slice the gDNA band from the gel and purify with a gel DNA extraction kit.
Hello Every one.! my down stream is to get rid of RNA from lysate and get DNA in the supernatant.
LOOKING FOR A PROCEDURE which WILL NOT use the below
RNase
phenol chloroform
Trizol
High centrifuge spins
Spin columns
Enzymes.
I have gone through few articles quoting the use of High concentration of KOH and MgCl2 at Ph >9 which will get the RNA denature due to transesterification.
Did any one try such chemical treatment procedures.
LiCl is known to selectively precipitate RNA.
https://www.thermofisher.com/se/en/home/references/ambion-tech-support/rna-isolation/general-articles/the-use-of-licl-precipitation-for-rna-purification.html
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