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RNase Protection Assay #1 Author: David Bowtell Source:University of Melbourne. Australia Abstract: Restriction digest 10 μg of plasmid DNA to linearize the plasmid
To the restriction digest add an equal volume of 2X PK buffer.
Incubate for 30 min at 37°C.
Add an equal volume of Phenol:Chloroform. mix well. and microcentrifuge for 3 min at full speed to separate the aqueous and organic phases.
Transfer the aqueous phase (upper phase) to a new tube and add to it an equal volume of 100% Ethanol. mix well. and microcentrifuge for 5 min at full speed to pellet the RNA.
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