Electrophoresis and its application

Electrophoresis:
Electrophoresis is separation technique based on movement of charge particle in an electric field.
Movement of charge particles can be determined by following formula--
V= Eq/f
Where,
V= Velocity of the charged particle;
E= electric field of the molecule;
q= Net charge of the molecule; and
f= Frictional co-efficient of the molecule
Types of electrophoresis:
1. Agarose gel electrophoresis ;
2. Poly-acryl amide gel electrophoresis [PAGE];
3. Sodium do-decyl sulphate Poly- acrylamide gel electrophoresis [SDS-PAGE] ;
4. Two dimensional –Poly-acrylamide gel electrophoresis [2D-PAGE];
5. Pulse field gel electrophoresis [PFGE];
6. Capillary gel electrophoresis [CGE]; and
7. Disc electrophoresis for Protein.

Application of electrophoresis:
1. Estimation of the DNA molecule.[ Agarose , PAGE ]
2. Analysis of PCR product. [ Agarose ]
3. Separation of restricted genomic DNA and RNA. [Agarose and PAGE respectively]
4. Conformation of newly isolated DNA .[Agarose]
5. Separation of most small fragments of DNA. [PAGE]
6. In forensic science.[Agarose , PAGE, SDS-PAGE, 2D PAGE ,Capillary gel electrophoresis , PFGE]
8. In determining molecular wt. of protein.[SDS-PAGE].etc

1. electrophoresis Presented By, Dipankar Saikia, M.Sc. in ZOOLOGY, UID-14PGZ10, DEPTT OF ZOOLOGY DARRANG COLLEGE

2. WHAT IS ELECTROPHORESIS Electrophoresis is separation technique based on movement of charge particle in an electric field. Movement of charge particles can be determined by following formula--- Where, V= Velocity of the charged particle; E= electric field of the molecule; q= Net charge of the molecule; and f= Frictional co-efficient of the molecule . V=E q ∕ f

3. types of electrophoresis 1 . Agarose gel electrophoresis ; 2. Poly-acryl amide gel electrophoresis[PAGE]; 3. Sodium do-decyl sulphate Poly- acrylamide gel electrophoresis[SDS-PAGE] ; 4. Two dimensional –Poly-acrylamide gel electrophoresis[2D-PAGE]; 5. Pulse field gel electrophoresis[PFGE]; 6. Capillary gel electrophoresis[CGE]; and 7. Disc electrophoresis for Protein.

4. Agarose gel electrophoresis PRINCIPLE: 1.It is a horizontal gel electrophoresis. 2.Molecules are separated on the basis of size , shape and charge. 3.To visualize movement of particle loading dye is added.

5. Fig: Agarose gel electrophoresis. source: https://www.bing.com/image

6. Poly- acrylamide gel electrophoresis [page] PRINCIPLE: 1.It is a vertical gel electrophoresis. 2.Negatively charged (-ve ) DNA and RNA molecule can be separated. 3.Molecules are separated on the basis of size , shapes and molecular weight.

7. Fig: Poly- acryl amide gel electrophoresis. [PAGE]. Source: http://www.google.com/image

8. SODIUM DO-DECYL –POLYACRYL AMIDE GEL ELECTROPHOERESIS[SDS-PAGE] PRINCIPLE: 1.Negatively charged Protein sample can be separated. 2.SDS detergent is used to give negative charge to the positively charged protein molecule. 3.Hydrocarbon tail of the SDS molecule is bind with the hydrophobic region of the protein molecule.

9. Fig:- SDS PAG source: https://www.bing.com/image E

10. TWO DIMENSIONAL POLY-ACRYL GEL ELECTROPHORESIS[2D-PAGE [2D-PAGE] PRINCIPLE: 1. It is a separation technique for Protein. 2. The proteins which have same molecular weight and iso-electric point can be separate. 3. The first dimension of it is vertical and second dimension is normal PAGE.

11. Fig:_2-dimensional Poly-acryl amide Gel Electrophoresis (2D-PAGE) . Source: http://www.google.com/image

12. PULSE FIELD GEL ELECTROPHORESIS [PFGE] PRINCIPLE: 1. Negatively charged large DNA molecule can be separated. 2. Above the length of 30-50 Kb molecules can be separated. 3. Here field direction is different.

13. Fig-Pulse field gel Electrophoresis. Source: http://www.google.com/image

14. CAPILLARY GEL ELECTROPHORESIS[CGE] PRINCIPLE: 1.It vertical electrophoresis. 2.Besed on shape, size, molecular weight, and electric charge.

15. Fig:- Capillary Gel Electrophores Source: http://www.google.com/image is.

16. DISCONTINUOUS[DISC] ELECTROPHRESIS PRINCIPLE: 1.It is a protein electrophoresis. 2.It solve the problem of aggregation and precipitation of protein. 3. Discontinuity is based on – *the gel structure; *the pH value and ionic streangth of the buffer; *the nature of the ion in the and the electrode buffer.

17. Fig: Discontinuous gel electrophoresis. source: https://www.bing.com/image

18. Application of electrophoresis 1. Estimation of the DNA molecule.[ Agarose , PAGE ] 2. Analysis of PCR product.[ Agarose ] 3. Separation of restricted genomic DNA and RNA.[Agarose and PAGE respectively] 4. Conformation of newly isolated DNA .[Agarose] 5. Separation of most small fragments of DNA.[PAGE] 6. In forensic science.[Agarose , PAGE, SDS-PAGE, 2D PAGE ,Capillary gel electrophoresis , PFGE] 8. In determining molecular wt. of protein.[SDS-PAGE].etc

19. references Dubey R .C ., 2013 a text book of Bio-technology. Reddy P. R.2012 Gel electrophoresis and its application(review paper)

20. THANK YOU