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Need to purify total RNA sample(contaminated with DNA)


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4 replies to this topic

#1 anonymous

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Posted 04 October 2001 - 09:00 PM

I have some RNA samples that I wish to use for RT-PCR that are contaminated with genomic DNA (I believe). Is there something I can do to remove the contaminating DNA from the samples.

Thanks in advance

#2 anonymous

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Posted 04 October 2001 - 09:00 PM

Ambion sells a product called DNA free. It's RNase free DNase with a DNase removal reagent - add - spin - done.

Eric


#3 anonymous

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Posted 04 October 2001 - 09:00 PM

Use RNase free DNase. Add this to the samples, incubate, and then re-precipitate the RNA to get rid of the enzyme. maniatis or current protocols probably has the details (concentration, duration, temp., RNA precipitation, etc)

#4 anonymous

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Posted 04 October 2001 - 09:00 PM

Dear Sir/Madam,

I routinely use acidic phenol (SIGMA, pH 4.3, Catolog No.: P 4682) extraction, because DNA is unstable below pH 7.0.

A more reliable method is that you can use DNase I (Gibco BRL, Catolog No.: 18068) Didestion.


#5 anonymous

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Posted 05 October 2001 - 09:00 PM

Use RNAse free DNAse (Sigma)




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