ELECTROPHORESIS

Presented By,Dipankar Saikia,

M.Sc. in ZOOLOGY,UID-14PGZ10,

DEPTT OF ZOOLOGY DARRANG COLLEGE

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

TYPES OF ELECTROPHORESIS1. 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]; and7. Disc electrophoresis for Protein.

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.

Fig: Agarose gel electrophoresis.

source: https://www.bing.com/image

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.

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

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.

Fig:- SDS PAGsource: https://www.bing.com/image

E

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.

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

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.

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

CAPILLARY GEL ELECTROPHORESIS[CGE]

PRINCIPLE:1.It vertical electrophoresis.2.Besed on shape, size, molecular weight, and

electric charge.

Fig:- Capillary Gel ElectrophoresSource: http://www.google.com/image

is.

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.

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

APPLICATION OF ELECTROPHORESIS1. 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

REFERENCESDubey R.C., 2013 a text book of Bio-

technology.

Reddy P. R.2012 Gel electrophoresis and its application(review paper)

THANK YOU