How to estimate content of RNA in a given sample by orcinol method

Estimation of RNA by Orcinol Method
To estimate RNA in a given sample by orcinol method.

When a solution of RNA is treated with orcinol. Reagent, it gets hydrolyzed to component base, phosphate and rebase reduce. Further heating with orcinol produces bluish green complex in presence of Fe+3.

Requirement: - Std. soln of RNA [0.05 mg/ml] sample solution of RNA, fresh orcinol reagent.

Procedure: - Mix 2 ml of sample and standard solution of RNA with 3 ml of orcinol reagent. Heat on boiling water bath for 30 mins. Cool and determine extinction at 665 nm. Against oreinol blank.

Calculation: - Conc. of sample = (Conc. Of standard / Absorbance of std.)x Absorbance of sample
                              

= 0.05 /0.249* 0.448


= 0.8945 mg / ml.

The conc. of RNA in given sample was found to be 0.8945 mg/ml.

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