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Gregory Wagner

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BIOL207 Lec22 20140305CHAPTER 8 contdA electrophoresis separation of molecules based on size httpwwwyoutubecomwatchvQEG8dz7cbnYFor routine DNA analysis agarose polysaccharide from seaweed is used most often a agarose is melted in electrophoresis buffer salts pH buffer then formed in a mold casting tray and allowed to cool b agarose gel is a 3D network of long polysaccharide molecules c DNA is applied to one end of the gel using a loading buffer eg glycerolvisible dye d Current applied across gel DNA move towardspositive electrodee Shorter molecules find their way through the gel faster than longer molecules so DNA is separated according to its length f DNA is visualized using a dye such as EtBr ethidium bromide which is fluorescent in UV light when bound to DNA g Need millions and millions of copies of a DNA molecule in order to detect it on an agarose gel with EtBr h Other types of gels are also used eg polyacrylamide for higher resolution in DNA sequencingB Blotting httpwwwyoutubecomwatchvPDxoNl7lRJka Some samples contain many molecules of different lengtheg a restriction digest of genomic DNA or an extract of mRNAb These will produce a smear rather than distinct ban
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