Then, vacuum pressure is provided in the container.Similarly, place the 3 MM Whatman paper above the membrane.After that, place the cut and wet nylon membrane on top of the gel.Then, position the agarose gel in such a way that the separated RNA faces above.Afterwards, place the gel above a sponge for support in a vacuum container containing SSC (transfer buffer).At first, cut a nylon membrane and 3 MM Whatman paper in the same size as denaturation gel.At last, run the gel at 125V for approximately 3 hours.After that, mix the RNA sample mixture in the equilibrated gel.Then, add RNA marker to the mixture above and incubate.Simultaneously, mix the sample with RNA loading buffer.After that, run the running buffer for equilibration for about 30 minutes.Firstly, prepare an agarose gel and pour it into a casting tray.DNA template Procedure of Northern Blotting RNA Extraction and Separation.Nylon hybridization membrane (3MM Whatman paper).Consequently, the detection of the desired RNA is done by using the radiolabeled probe with all or a part of the base sequence of the targeted RNA hybridizing with the immobilized and separated RNA.The principle of the method is the separation of RNA according to size by agarose gel electrophoresis and afterwards transferring the separated RNA on a nylon membrane.And also determine what the regulating factors of the gene expression are.Identify the types of protein secreted by specific tissues.In short, Northern blotting is the process of measuring or quantifying RNA, especially mRNA. Like all the blotting techniques, northern blotting is the process of blotting separated RNA in a membrane, hybridizing it with the probe, and quantifying the RNA. Many techniques help in determining the type of protein and the reason for its expression. A person with a genetic mutation expresses a new or foreign protein that may or may not be harmful.
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