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Northern BlottingAdapted from Molecular Cloning (Maniatis), Current Protocols in Molecular Biology, and J. Yost Lab (UMN)
Objective:Northern blotting allows detection of specific RNA sequences. RNA is fractionated by agarose gel electorphoresis, followed by transfer (blotting) to a membrane support, followed by hybridization with DNA or RNA probes.
Procedures:
Start=> This protocol requires isolated total RNA or poly(A)+ RNA.
- You must first run the RNA on a denaturing gel; a formaldehyde or glyoxal/DMSO gel.
- Next, you need to transfer the RNA from the gel to a solid support membrane, preferably nylon.
- Once, the membrane with the immobilized RNA is available, hybridization with the appropriate probe is required.
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Northern Blots