Gel Electrophoresis

A tool for qualitative estimation of DNA/RNA

• gel electrophoresis is a powerful and widely used method that
separates molecules on the basis of electrical charge, size, and shape.
The method is particularly useful in separating charged biologically
important molecules such as DNA (deoxyribonucleic acids), RNA
(ribonucleic acids), and proteins. Agarose forms a gel like consistency
when boiled and cooled in a suitable buffer.
• Making a 1% Agarose Gel (for DNA sample)
• Loading of Samples
• Examining the gel
• Precautions:
• Wear gloves during the addition of EtBr and while handling the casted
gel (EtBr is a potent carcinogen).
• Handling the gel should be careful as the gel may break due to
improper handling.
• While performing the UV-trans illumination for visualising the bands,
avoid direct contact and exposure to eyes.
• RNA has the tendency form both secondary & tertiary structure by
varied intra-molecular base pairing which impedes separation by
electrophoresis. Consequently electrophoresis in RNA is performed in
denatured condition.
• Preparation of gel
• Loading of Samples
• Examining the gel
• Connect the power cord to the electrophoretic power supply according to the
convections: Red-anode and Black-cathode.
• Electrophoreses at 100 volts (this may change) and 70mA until dye markers
have migrated an appropriate distance, depending on the size of RNA to be
visualized
Application
• Verifying molecular markers
• Verifying gene expression
• In medical science
• In forensic science
• Checking quality & quantity of DNA