Separate & Transfer Your RNA Step 1: Gel. With basic precautions, such as cleaning the mold from your workbench and wearing gloves, isolating and maintaining the integrity of your RNA is easy. So don’t let the idea of working with RNA scare you. There are numerous RNA isolation kits on the market now, and lots of RNAse inhibitors available. I think that Northern blots got a bad rap from the days when working with RNA was as hard as it got in molecular biology. And the naming tradition just continued with Western blots. So when invented the RNA method was named “Northern blot” as an homage to the original nucleic acid detection method. Northern Blots are done the same way as Southern blots but RNA is detected instead of DNA. A typical Southern blot experiment goes as follows: 1) run a DNA gel, 2) transfer gel contents onto a membrane, 3) hybridize said membrane with radioactive DNA probe, 4) wash off unbound radioactive probe, and 5) detect radioactive probe. Southern blots are a method of detecting DNA and named after the surname of their inventor Ed Southern. Northern Blots are named after their big brother: The Southern blot.
#Northern blot analysis of rna how to
However, because of its unpopular nature finding someone that can teach you about how to do a good Northern Blot can be hard. So sometimes Northern Blots are a necessary evil. Yet, qRT-PCR is prone to false positives and negatives, and reviewers may require Northern Blot confirmation of your qRT-PCR results. Northern Blots fell out of favor for two reasons: The perceived difficulty of working with RNA and because most people don’t like working with radioactivity. After all, it is more common these days to detect and quantify RNA with quantitative real time (qRT)-PCR than with Northern Blots. You might think Northern Blots are an old-fashioned technique.
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