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Agarose Gel Electrophoresis Video Lecture | Crash Course for CSIR NET Life Sciences

FAQs on Agarose Gel Electrophoresis Video Lecture - Crash Course for CSIR NET Life Sciences

1. What is agarose gel electrophoresis and how does it work?

Ans.Agarose gel electrophoresis is a laboratory technique used to separate nucleic acids (DNA or RNA) based on their size. The process involves preparing an agarose gel, which is a gelatinous substance made from seaweed. The gel is cast into a mold and allowed to solidify, creating a matrix through which molecules can migrate. When an electric current is applied, negatively charged nucleic acids move towards the positive electrode. Smaller fragments travel faster and further through the gel than larger ones, allowing for size-based separation. After electrophoresis, the separated bands can be visualized using staining agents like ethidium bromide or SYBR Green.

2. What are the components required for agarose gel electrophoresis?

Ans.The key components required for agarose gel electrophoresis include agarose powder, a buffer solution (commonly TAE or TBE buffer), DNA samples, a loading dye, a gel casting tray, and an electrophoresis chamber. Agarose powder is mixed with the buffer and heated to create the gel. The gel is then poured into a casting tray where it solidifies. The DNA samples, mixed with loading dye for visibility, are loaded into wells created in the gel. The electrophoresis chamber provides the necessary electric field to facilitate the movement of the DNA through the gel.

3. How do you prepare an agarose gel for electrophoresis?

Ans.To prepare an agarose gel, first, determine the desired gel concentration based on the size of the DNA fragments to be separated (typically between 0.7% and 2%). Measure the appropriate amount of agarose powder and buffer solution. Heat the buffer on a hot plate until it boils, then add the agarose powder and stir until fully dissolved. Allow the solution to cool slightly before pouring it into a gel casting tray with a comb set in place to form wells. Once the gel has solidified, remove the comb and place the gel in the electrophoresis chamber filled with buffer.

4. What are some common applications of agarose gel electrophoresis?

Ans.Agarose gel electrophoresis is widely used in molecular biology for various applications, including DNA fragment analysis, restriction fragment length polymorphism (RFLP) analysis, polymerase chain reaction (PCR) product verification, and cloning procedures. It is also utilized in the analysis of RNA for quality control in experiments and in forensic science for DNA profiling. The ability to visualize and quantify DNA or RNA fragments makes it an essential tool in genetic research, diagnostics, and biotechnology.

5. What safety precautions should be taken while performing agarose gel electrophoresis?

Ans.When performing agarose gel electrophoresis, several safety precautions should be followed. First, always wear gloves and safety goggles to protect against exposure to hazardous chemicals, such as ethidium bromide, which is a mutagen. Additionally, ensure that the electrophoresis chamber is handled with care to avoid electrical hazards. Dispose of any waste, including used gels and staining solutions, according to local regulations. It is also important to work in a well-ventilated area and follow the manufacturer's instructions for all reagents and equipment used in the process.

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Sep 28, 2025 Last updated

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