Agrose gel electrophoresis techniques.

1. M.Sc
Micribiology
1st Year
“Agarose Gel Electrophoresis”
By
Vivek Kumar
Bangalore
university

8. An agarose gel is
prepared by
combining agarose
powder and a buffer
solution.
+
Making an Agarose Gel

9. Buffer System
• Various buffer are used for agarose electrophorosis.
• Most common is 1) TAE (Tris,Acetate.EDTA)
2) TBE (Tris, Borate, EDTA)
• Buffers not only establish an ideal pH , but provide
ions to support conductivity.
• Buffer maintains the pH.

10. Add enough electrophoresis buffer to cover the gel.
Make sure each wells is filled with buffer.
Place the gel in the electrophoresis chamber

11. Loading the Gel
Carefully place the pipette tip over a well and gently expel the
sample.
The sample should sink into well.
Be careful not to puncture the gel with the pipette.

12. Running the Gel
Place the cover on the gel electrophoresis chamber.
connecting the electrical leads to the power supply.
Be sure the leads are connected correctly.
DNA migrates towards the anode.
When the power is turned on ,bubbles should form on the
electrods in the electrophoresis chamber.

18. Application
•Estimation of the size of DNA molecules.
•Analysis of products of a polymerase chain
reaction (PCR).
•Separation of DNA fragments for extraction
and purification.
•In the field of forensic science, molecular
biology.

19. Advantages
• The main benefit of agarose gel technique is that it can be
easily processed.
•Agarose gel does not denature the DNA samples and they
stay in their own form.
Disadvantage
•There is also a disadvantage of gel electrophrosis
that it may melt when the electric current is passed
through it.

20. Thank you