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Cannot get ride of gDNA before ReverseT PCR

H.M.Romero-Arce at hgmp.mrc.ac.uk H.M.Romero-Arce at hgmp.mrc.ac.uk
Thu May 15 14:15:34 EST 2003


Hello, hope someone can give me some advice.

I did DNA/RNA soil extraction following Griffitts, 2000 method of 
extraction. 
Then I stored the extracted DNA/RNA in 1x TE buffer. My problem is that now 
that I want to digest the DNA to get only the RNA for RT-16SPCR it's 
impossible to digest completely the DNA. I read that there is a limited 
amount 
of DNA that can be digested in the DNase assay. To start with, my samples 
got 
a lot of DNA, they were stored in a buffer containing EDTA and I cannot 
modify 
the protocol for extraction because by now population in the soil has 
changed 
so I need to work with these extracts. Dilution is not an option because 
that 
would cause the lost of RNA which belongs to minority species in the soil.

I'd like to know if a part from the many kits that are in the market to 
purify 
RNA from gDNA there is a 'lab-made' way of doing so, taking in account the 
state of my extracts.

Thank you.

Monica




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