RNase Protection Assay (RPA) for quntifying Gene Expression Levels

RNase protection assay (RPA) is one of the most common and standard techniques that has been used for analysing the level of gene expression by quantifying mRNA molecules in the cell. This technique provides more sensitivity and reproducibility for measuring mRNA more than the conventional approaches such as Northern blotting. The principle of the technique is based on the hybridization of the target mRNA to a radioactively labelled RNA probe. The hybridized part of the complex becomes protected whereas the unhybridized part of the RNA probe is digested with specific ribonuclease. The hybrid can be resolved by a denaturing gel. Subsequent detection will reveal the appropriate-sized gel band corresponding to the target mRNA. The major advantage of RPA over the other techniques is that the expression of up to 10 or 12 mRNAs can be studied simultaneously in a given sample.

Reference:
http://www.springerlink.com/content/r413643m921186m6/fulltext.pdf

Amjad yousuf
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