12/6/2023 0 Comments Application of southern blotting![]() ![]() How does the DNA actually go from the gel into the blot? Even after leaving the blotting paper overnight, the transfer may not be complete! As you can imagine, the larger a piece of DNA is, the slower it will migrate through an agarose gel. It’s because these large pieces of DNA don’t transfer very well to the blotting paper. Why is there is a second denaturing step for DNA that’s 15 KB or larger? The steps for this process are described in the following image: Finally, this “probed” radioactive blot is then imaged using an autoradiograph. After the “blotting” process is complete, the DNA is then “probed” using a radioactive (or fluorescent) DNA sequence that is complementary to the sequence that you want to detect. Next, in a similar fashion to Western blots, this DNA is then transferred onto blotting paper (typically made of nitrocellulose or nylon). This will separate the DNA by size and you’ll know which well in the gel corresponds with which sample. Next, digest the DNA using DNAse and run these fragments on an agarose gel. This can be from a cell lysate or from tissue samples, etc. ![]() To execute a southern blot, first collect some DNA. As you probably understand, this is one of the most common techniques that geneticists and molecular biologists know because everything they do needs to be proved by a southern blot. You can also do some amazing things like making transgenic mice and proving that you have selected for certain genes of interest to you! All you have to do is Southern Blot any set of DNA from the mice and you’ll have concrete proof. Most commonly, you will be testing for DNA that you get from cell lysates or after creating your own plasmids. Southern Blotting is a technique that is used to detect whether you have a specific DNA sequence available in your sample. This method was originally published on here: Southern Blot Scientific Method. To isolate and identify desire gene of interest.Search Engine for Scientific Methods.Used for paternity testing, criminal identification, victim identification. ![]()
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