The nuclease protection assay is a method used in biochemistry
and genetics for the detection, quantitation and mapping of specific RNAs in a
complex mixture of total cellular RNA. This technique includes both
ribonuclease protection assays (RPAs) and S1 nuclease assays.
The principles of this technique are the
hybridization of single-stranded RNA. This is achieved by addition of antisense
RNA probe, containing the complementary strands of RNA, to the initial solution
containing the RNA. After hybridization, the remaining RNA, which did not
hybridize, is removed by adding nuclease. Then, the existing
nucleases in the solution are inactivated and, at the same time, the hybridized
RNA precipitates. These are separated on denaturing polyacrylamide gel and visualized by autoradiography.
Pl 1 – Grupo 3
Ana Catarina Azevedo
Bruna Martins
Eliana Santos
Marta Monteiro
Ricardo Rosa
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