Electrophoresis & Multiple Myeloma Lecture 2017

  • Electrophoresis is the motion of dispersed particles in a fluid (water) under the influence of constant electric field.
  • Consist of: gel, samples, buffer (to supply ions) and electrical field (cathode & anode & battery & wires).
  • Cathode (-) reduction takes place and Anode (+) oxidation take place
  • Technique requires a dye for coloring the sample.
  • Usually used in Biochemistry lab to separate DNA, RNA, and proteins.
  • Thisphenomenon was observed for the first time in 1807 by Ferdinand Frederic Reuss (Moscow State University).
  • This method is the basis for a number of analytical techniques used in chemistry for separating molecules by size, charge, shape or binding affinity.
  • Electrophoresis of positively charged particles (cations) is calledcataphoresis, while electrophoresis of negatively charged particles (anions) is called anaphoresis.
  • Electrophoresis is a technique used in laboratories in order to separate macromolecules based on size.
  • The technique “applies a negative charge” so proteins move towards a positive charge anode. This is used for both DNA and RNA analysis. Similar to the Sebia analyzer method for SPE/IFE & UPE/UIFE.
  • Polyacrylamide gel electrophoresis(PAGE) has a clearer resolution than agarose and is more suitable for quantitative analysis.
  • We can use DNA ladder (bp)/Calibrator or IQC samples (normal & abnormal) for quality assurance purpose.
  • Agarose gel for very big macromolecules separation. Have very big pore sizes. PAGE used for small molecules in sizes because it has small pores in size.

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