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Southern blot e northern blot12/30/2023 RNA separation: RNA is isolated from the source by cell lysis and removing DNA and protein contaminants using enzymes such as protease and DNAse.The steps involved in the blotting procedure include: The technique was developed by Alwin in 1979 to study known mRNA sequences. The technique is used to detect and analyze specific RNA molecules from a mixed RNA sample. Visualization: For the visualization of separated bands, a dye such as EtBr (ethidium bromide) is added to the DNA sample so the fragment glows when observed under UV light.įigure: Southern blotting procedure.Hybridization: A labeled hybridization probe (a DNA fragment with a homologous sequence to the target DNA) is used to bind to the membrane to initiate a measurable reaction. Transfer: The DNA bands are transferred on a nylon or nitrocellulose membrane and made permanent by exposure to UV rays.
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