Basic Protocol For Northern Blotting

Northern blotting is the transfer of RNA from a gel to a filter, and the method is comparable to that for Southern blotting with only a few modifications. By adding formaldehyde to the agarose gel, the RNA is denatured and secondary structures are prevented. Soak the gel in transfer buffer to dilute out the formaldehyde, cut off the marker lane for ethidium bromide staining, but do not stain the main body of the gel because this reduces RNA transfer.[13] Proceed as described earlier, although denaturing and neutralizing treatments are not essential if the RNA fragment is less than 5 kb. After the overnight transfer, the RNA can be cross-linked to the filter by baking or UV treatment prior to hybridization.

When compared to Southern blots, Northern blots, because of the ubiquitous nature of RNases, are highly variable in terms of sensitivity. As the presence of RNases is common, it is important to ensure all solutions are diethyl pyrocarbonate (DEPC)-treated, all plastics are baked prior to use, and, if possible, individual apparatus should be provided for RNA use. These preparations may help to provide more consistent results.

Getting Started With Dumbbells

Getting Started With Dumbbells

The use of dumbbells gives you a much more comprehensive strengthening effect because the workout engages your stabilizer muscles, in addition to the muscle you may be pin-pointing. Without all of the belts and artificial stabilizers of a machine, you also engage your core muscles, which are your body's natural stabilizers.

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