DNAse 1 treatment of RNA samples

John F Mackay MackayJF at bmnz.co.nz
Thu Nov 16 19:23:34 EST 2000


Hi,

DNas'ing RNA samples regularly makes the pages of BioTechniques (and this
newsgroup). The latest ones seem to advocate some, one or all of the
following:

- 1-2U DNase I per ug RNA

- Buffer containing Mg and Mn. Buffer should be lower than 7 or higher than
9. Also useful to contain 0.1mM DTT

- 10 minutes at 37 degrees

- Add EDTA to 2.5mM final concentration

- heat-kill at 75 degrees for 5 minutes. Not killed if standard PCR buffer
is used ie. pH 7-9 (but this paper didn't use the EDTA - which was the
subject of another paper!)

The above is puled from about 4 different papers in BioTechniques. Searching
their site under 'DNase' should pull them out. . . . .



Cheers,

John


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