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Northern Analysis: The Basics

discussion of this subject, see "Technical Bulletin 169: Membrane Transfer and Crosslinking for RNA."

Incorporated into the NorthernMax and NorthernMax-Gly procedure is a rapid, alkaline transfer method that increases blot sensitivity by efficiently moving RNA, especially larger transcripts, onto the membrane. This step only takes 2 hours and can trim an almost entire day off the standard procedure, which generally requires overnight transfer. The NorthernMax and NorthernMax-Gly Kits have been optimized to work with Ambion's BrightStar-Plus positively charged nylon membranes, and we recommend their use to minimize background and maximize signal. Please note that nitrocellulose membranes are chemically incompatible with the NorthernMax Transfer Buffer, and should not be used with these kits.

Once the RNA is transferred, the membrane should be immediately treated to crosslink the RNA. This can be done by ultraviolet light (the preferred method) or by baking. For more information, see "Technical Bulletin 169: Membrane Transfer and Crosslinking for RNA."


Prehybridization and Hybridization with Probe

Prehybridization, or blocking, is required prior to probe hybridization to prevent the probe
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