RNA was mixed with 0.1 vols of 3M NAOAc (pH = 5.2) and 2.5 vols of 100% EtOH, vortexed and incubated @ -20C for 30mins. RNA was pelleted @ 16,000g, 30mins, 4C. Supe was discarded and pellet was washed with 1mL 70% EtOH. RNA was pelleted @ 16,000g, 15mins, 4C. This was step was repeated a second time. Supe was discarded, the RNA was resuspended in 50uL of 0.1% DEPC-H2O, spec’d and stored @ -80C:
Results:
Interestingly, precipitating the samples vastly improved the 260/230 ratios. However, the 260/280 ratios DECREASED for all samples except one (CA 1). Not really thrilled about this fact, nor am I sure why this would happen.
RNA was stored @ -80C in “Sam’s RNA Box #1.”