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BIOC23H3 Lecture : Lec 7 - Gel electrophoresis.doc

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School
UTSC
Department
Biological Sciences
Course
BIOC23H3
Professor
Rongmin Zhao

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Document Summary

Low voltage thin sheet electrophoresis: supporting medium, 1) paper. Made of cellulose; hydrophilic; absorb water and mocs can separate on paper. Only low voltage can be used; can"t readily adjust porosity. Suitable for microtechniques such as separation involving immunodiffusion and immunoelectrophoresis. Better resolution than paper but still some diffusion. Can"t easily preserve separating mocs: 3) thin layer of material = thin layer electrophoresis. Plates placed horizontally on electrophoresis units and thin layer allowed to saturate w/ buffer by diffusion from reservoir. Similar to tlc (but tlc moves moc by organic mobile phase whereas tle moves mocs by electric field: gels: Jello-like semi-solid matrix heat agarose in appropriate buffer 80-100 homogenous gel matrix. Pore size = large good for separating large mocs like. Dna, rna (which are bigger than proteins: polyacrylamide. Polya gels formed by copolymerization of acrylamide (water soluble monomer) + cross-linking agent bis. Polymerization done by chemical reaction using ammonium persulfate (initiator) and quaternary amine, temed = catalyst.

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Related Questions

Agarose gel electrophoresis is used to separate charged molecules. Why do nucleic acids migrate from the cathode (-ve electrode) to the anode (+ve electrode) during agarose gel electrophoresis?

BACKGROUND INFO:
For purposes of genetic counseling, you wish to determine thehemoglobin genotypes for a couple who both have family histories ofsickle cell disease.
Since hemoglobin S - Hb S - and hemoglobin A - Hb A - havedifferent net charges, if each is placed in an electrical field,the two types of hemoglobin proteins will migrate differently.(Note: Separation of molecules via this type of electrophoresis -unlike the gel electrophoresis considered in Unit 16 - depends onthe different net electrical charge (not the size – which isessentially the same for both hemoglobin proteins.)

QUESTION:
Will Hb A or Hb S move further toward the positive pole uponelectrophoresis? Hint: Recall the difference in charge due to thepoint mutation substituting valine for glutamate in Hb S.

Gel electrophoresis is a process used in laboratories to separate macro molecules. Mark the statements that are true as related to gel electrophoresis. An electrical charge is applied to the gel plate The pieces of DNA move due to electrical charge and also size A sample of the DNA is placed in a small well and a charge is applied The pieces of DNA or bands are stained to be visible Running a "gel" can be used to compare a perpetrators DNA to the DNA found at a crime scene.