Estimation of RNA in unknown sample using Orcinol Reagent • Label 1 to 7 test tubes as Blank, 01, 02, 03, 04, 05 and Unknown sample. • Pipette out different dilutions of Standard RNA solution (500µg/mL) in the test tubes. • Make final volume as 2.0 mL using 5% Perchloric acid. • In the test tube labelled as Blank, pipette out 2.0 mL of 5% Perchloric acid. • Pipette out 2.0 mL of Unknown sample in the last test tube. • Add 3.0 mL of Orcinol Reagent to all the tubes. • Cover the tubes with Foil paper. • Keep the tubes in boiling water bath for 15 minutes. • Remove the tubes and let them cool. • Record the absorbance at 665 nm using Blank as Reagent blank. • Draw a standard curve of Absorbance v/s RNA concentration. • From the absorbance of Unknown sample, determine the amount of RNA in it.

• published: 18 Feb 2022
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