Checking+integrity+of+RNA

Incubate one aliquot of 250-1000 ng of RNA at -80 C and one aliquot at 37 C. Perform electropheresis with these aliquots on a 0.8% gel with a 1 kb ladder. Run the gel at 220 V for 20 min. If the RNA solution does not contain RNases or other contamination then the 37 C sample should be about as bright as the -80 C sample. Also, if the sample incubated at 37 C shows a lower 28S:18S ratio than the control, the RNA may have contaminants. Two prominent bands should be visible: 28S band around 4.5 kb and 18S band at 1.9 kb. The intensity ratio of the 28S:18S band should be 1.5-2.5:1.

- I originally had this note below in the protocol, but it does not seem to be accurate based on my experience from 2-24-13 in notebook. Run the gel at 70 V or another voltage, but not too high. Do not use high voltage in an attempt to avoid RNA degradation during electrophoresis (http://www.evrogen.com/technologies/RNA-electrophoresis.shtml) <- this does not seem to be accurate