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Page Title:
Determining RNA purity and yield |
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 ERDC TN-DOER-R4
September 2004
20. Determine RNA purity and yield.
21. Adjust RNA concentration to 2 g/L with RNase-free water or with RNA storage solution
for long-term storage.
22. Store RNA in 25 L (50 g) aliquots at -80C.
Determining RNA purity and yield:
Materials needed
10 L pipette and tips
RNA storage solution (0.1 mM EDTA)
1000 L pipette and tips
O.D. buffer (50 mM Tris, 0.1 mM EDTA, pH 7.5)
1-ml quartz cuvettes (2)
1.
Transfer 4 L of RNA sample into a 1-mL quartz cuvette.
2.
Add 996 L of O.D. buffer.
3.
Measure A260 and A280 using 1-mL O.D. buffer as a blank. Calculate yield:
Total A260 = A260 of dilute sample x 250 (dilution factor)
Concentration (g/mL) = Total A260 x 40 g/L
Yield (g) = volume of RNA (mL) x concentration (g/mL)
Purity = A260/A280
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