Electrophoresis & its Type

Electrophoresis & its Type

1. COURSE TITLE :-Basic Concept Laboratory techniques COURSE NO. :-PGS504 COURSE CREDIT :- 1(0+1) SUBIMTTED T0:- SUBMITTED BY:- Dr. Anurag sir Name :- Neelima singh College Id:- 20200300 Dept. :- PMBB Topic:- E lectrophoresis & it's type Indira Gandhi Krishi Vishwavidyalaya COLEGE OF AGRICULTURE, RAIPUR

2. Introduction Definition Electrophoresis apparatus Supporting media for electroforesis Working principle Classification Factors Affecting Advantages and disadvantages CONTENT

3. Introduction Father of electrophoresis: Arne Tiselius (Nobel Prize in 1948) Electrophoresis is a physical method of analysis which involves separation of the compounds that are capable of acquiring electric charge in conducting electrodes. Electrophoresis separates the  biomolecules  like proteins, nucleic acids, amino acids etc. according to their size, shape, chemical charge etc. Movement of positively charged ion towards negatively charged electrode refers to “ Cataphoresis ”. Movement of negatively charged ion towards the positively charged electrode refers to “ Anaphoresis ”.

4. Defination of Electrophoresis “ Electrophoresis may be defined as the migration of the charged particle through a solution under the influence of an external electrical field.” Supporting media Paper Gel Cellulose acetate

5. Electrophoresis Instrument

6. Type of Electrophoresis

7. Free Electrophoresis: In this type of electrophoresis a free electrolyte is taken in place of supporting media. It is mostly of two types micro-electrophoresis moving boundary electrophoresis mostly used in non-biological experiments. Free Flow Electrophoresis is an electrophoresis procedure working continuously in the absence of a stationary phase (or solid support material such as a gel).

8. Free Electrophoresis Micro Electrophoresis Moving Boundary Elecctrophoresis

9. Zone Electrophoresis: It involves the migration of the charged particle on the supporting media can be Paper, Cellulose acetate membrane, Starch Gel, Polyacrylamide . Components separated are distributed into discrete zone on the support media. Supporting media is saturated with buffer solution, small volume of the sample is applied as narrow band.

10. Paper Electrophoresis Used - separation of small charged molecules such as amino acids and small proteins Filter paper was the first anti-convection medium to be widely used The support medium is a filter paper

11. Gel electrophoresis Used - separation of DNA, RNA or proteins. Gel material acts as a "molecular sieve”. Gel is a colloid in a solid form (99% is water). Types of gels :- Agar and Agarose gel, Starch, Sephadex , Polyacrylamide gels.

12. Agarose gel electrophoresis Agarose  gel the supporting media in the electrophoresis. used - the electrophoresis of  DNA ,  RNA  and  Protein

13. Pulse field gel electrophoresis (PFGE ) This technique is invented by Schwartz and Cantor in 1984. used - separate large DNA molecule DNA fragments up to 10 mb can be separated by this technique