I extracted RNA from skin and quantified it in an Nanodrop machine. The 260/230 ratio was very low and two peaks appeared in the absorbance curve, but the RNA is ressuspended in RNAse free water. How can I clean it?
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$\begingroup$ wash it with chloroform.. $\endgroup$– WYSIWYGMay 7, 2013 at 10:29
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Phenol absorbs at 230 nm, this is the most likely contamination in your case. You should try to avoid pipetting from the phenol phase, or use a phase lock gel to make this easier. I assume you washed it a few times with pure chloroform after the phenol/chloroform mixture, that should help to get rid of the phenol.
I only precipitate my RNA with 1-1.5 volumes of isopropanol after the chloroform/phenol extraction, that should be sufficient for most uses. Just add the isopropanol, keep it at -20°C for 30 minutes and then centrifugate it at 9000 g for 20 minutes, redissolve the pellet in water.
answered May 7, 2013 at 10:30