Lab 4 - Electrophoresis

Lab 4 - Electrophoresis - Electrophoresis INTRODUCTION:...

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Electrophoresis INTRODUCTION: Electrophoresis is a method for separating macromolecules, by either nucleic acids or proteins based on size, charge, and/or physical properties. We will be conducting 1D-PAGE, 2D-PAGE, and isoelectric focusing. PRINCIPLE: Electrophoresis is a technique that causes the charged particle to move based on the mass and charge of the particles. The mass of the particle is important because the greater the mass the greater the mass of the particle thereby making the particle to migrate base on the charge-to-mass ratio. Some advantages of using Polyacrylamide Gel Electrophoresis is that it has a high resolving power for only using a small amount of protein and nucleic acid, acceptance of relatively large sample sizes, minimal interaction of migrating molecules with matrix, and physical stability of the matrix. 1 With different methods of electrophoresis, we can find more or less information about the proteins that we are testing. Using one dimensional polyacrylamide gel electrophoresis (1D-PAGE), we are limited in the information we can gain from the protein. 1D-PAGE only gives information about the molecular size of the particle. Using 1D-PAGE, we are limited because if two proteins have the same molecular size then the researcher does not know if there are multiple proteins or just the desired protein. The problem is resolved when doing 2D-PAGE; 2D-PAGE allows the researcher to view the protein or nucleic acid from the charge as well as molecular size. Even though two
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This note was uploaded on 01/26/2009 for the course BIOLOGY 344 taught by Professor Cheng during the Spring '05 term at Temple.

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Lab 4 - Electrophoresis - Electrophoresis INTRODUCTION:...

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