cloudy RNA preparation - (May/12/2006 )
I've just isolated some RNA from cells using Tri reagent & the final prep is cloudy. The ratios don't look too bad & it looks fine on a gel but we think it's either protein or salt precipitation.
Is there anything we can do to clean it up ? We're intenting to use it for cDNA synthesis for use on an ABI microfluidic card/low density array-will it be ok for this ???
Check the 260/230 ratio - Affy strongly suggest that RNA with ratio of less than 1.8 should not be applied to their arrays
I'm sure fred will give you some advice on cleaning up your Trizol prep, he is the king of Trizol after all
hi dixon ! if you can afford again give phenol: chloroform phase separation, precipitating the upper aqueos layer with chilled isopropanol, and redissolving the RNA after a 70% ethanol wash.
phenol: chloroform separation would remove protein contamination if any and isopropanol precipitation step would remove salts
all the best