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1. Techniques in Biotechnology…
M.PHOOL BADSHAH

2. Lecture-6
Agarose Gel Electrophoresis

3. Introduction
• The act of separating and studying macromolecules such as
DNA, RNA, or proteins based on their size is a widely used
technique in molecular biology, biotechnology, and
biochemistry.
• It is a seaweed-derived carbohydrate polymer that, when
coupled with a buffer and heated, forms a matrix resembling
gel.
• This gel is then used as a support medium for electrophoresis.
• Here is a general rundown of the procedure:

4. 1- Preparing the Gel:
 Agarose powder is mixed with a buffer solution and heated to
dissolve the powder.
 A melted agarose solution is poured into a gel mould.
 A comb is inserted into the gel to create wells for loading
samples.
2- Loading Samples:
 To give the samples colour and density, proteins, RNA, or DNA
are mixed with a loading buffer.
 The samples are progressively added to the wells of the agarose
gel.

6. 3- Running Electrophoresis:
 The gel is placed inside an electrophoresis chamber that has been filled
with buffer solution.
 When an electric current is passed across the gel, the charged
macromolecules move through the gel matrix.
 Smaller molecules travel farther and faster over the gel than larger ones
do.
4- Visualization:
 After electrophoresis, the gel is treated with a dye that binds to
proteins, RNA, or DNA and makes them visible.
 The gel is then subjected to UV light, which makes it possible to
see and record the separated bands.

9.  Note:
• Agarose gel electrophoresis performs several tasks, such as
size analysis of DNA fragments, confirmation of molecular
biology processes like polymerase chain reaction (PCR), and
assay purity verification of DNA or RNA materials.
• It is a crucial instrument for molecular biology research and
diagnosis.