ELECTROPHORESIS

Group A February 26th, 2018

Gel electrophoresis is a laboratory method

used to separate mixtures of DNA, RNA, or proteins
according to molecular size. In gel electrophoresis,
the molecules to be separated are pushed by an
electrical field through a gel that contains small pores.
The molecules travel through the pores in the gel at a
speed that is inversely related to their lengths. This
means that a small DNA molecule will travel a
greater distance through the gel than will a larger
DNA molecule.
The rate of migration of charged molecules
Types of electrophoresis are:
depends upon following factors: 1. Polyacrylamide Gel Electrophoresis (PAGE)
(a) The strength of electric field, size and shape. 2. Discontinuous Gel Electrophoresis
(b) Relative hydrophobicity of the sample. 3. Nucleic Acid Sequencing Gels
(c) Ionic strength and temperature of the buffer. 4. Sodium Dodecyl Sulfate- Polyacrylamide Gel
(d) Molecular size of the taken biomolecule. Electrophoresis (SDS-PAGE)
(e) Net charge density of the taken bio mole-cule. 5. Agarose Gel Electrophoresis
(f) Shape of the taken biomolecule. 6. Pulsed Field Gel Electrophoresis (PFGE)
As previously mentioned, gel electrophoresis 7. Isoelectric Focusing of Protein
involves an electrical field; in particular, this field is 8. Two-Dimensional Electrophoresis of Proteins
applied such that one end of the gel has a positive 9. Capillary Electrophoresis (CE)
charge and the other end has a negative charge. 10. Immunoelectrophoresis (IE)
Because DNA and RNA are negatively charged
molecules, they will be pulled toward the positively
charged end of the gel. Proteins, however, are not Discuss and answer the following questions with
negatively charged; thus, when researchers want to your group. Find the answers in these
separate proteins using gel electrophoresis, they must references, Boyer (2012) pages 165-186; Wilson
first mix the proteins with a detergent called sodium & Walker (2010) pages 399-419.
dodecyl sulfate. This treatment makes the proteins
unfold into a linear shape and coats them with a 1. What is the principle of electrophoresis?
2. What are the factors affecting molecular
negative charge, which allows them to migrate
mobility of electrophoresis?
toward the positive end of the gel and be separated.
3. Compare the types of electrophoresis, including
Finally, after the DNA, RNA, or protein molecules
the medium?
have been separated using gel electrophoresis, bands 4. Show how to calculate molecular size with
representing molecules of different sizes can be electrophoresis?
detected.
All modes of electrophoresis are based on the
principles just outlined. The major difference among
the various methods is the type of support medium.
Cellulose and cellulose acetate are used as a support
medium for low-molecular-weight biochemical like
amino acids and carbohydrates. Polyacrylamide and
agarose gels are widely used as support media for
larger molecules. In capillary electrophoresis, several
different types of support media, including the
natural, untreated surfaces inside a silica narrow bore
are used.
Electrophoresis