DNA fragments generated by the restriction endonucleases in a chemical...
DNA fragments generated by restriction endonucleases in a chemical reaction can be separated by gel electrophoresis. Since DNA fragments are negatively charged molecules they can be separated by forcing them to move towards the anode under an electric field through a medium/matrix. The DNA fragments separate according to their size through sieving effect provided by matrix.
View all questions of this test
DNA fragments generated by the restriction endonucleases in a chemical...
The cutting of DNA by restriction endonucleases results in the fragments of DNA nd these fragments can be separated by gel electrophoresis as DNA fragments are negatively charged molecules they can be separated by forcing them to move towards the anode under an electric field through a medium or matrix nd then these DNA fragments separate according to their size through sieving effect provided by agarose gel !!!
DNA fragments generated by the restriction endonucleases in a chemical...
Electrophoresis:
Electrophoresis is a technique used to separate DNA fragments based on their size and charge. It involves the movement of charged molecules under the influence of an electric field through a gel matrix. In the context of DNA fragments generated by restriction endonucleases, electrophoresis is the most appropriate method for their separation.
Principle of Electrophoresis:
When an electric field is applied to a gel matrix containing DNA fragments, the negatively charged DNA molecules move towards the positive electrode. The movement of DNA fragments through the gel is influenced by their size and shape. Smaller fragments move faster and migrate further through the gel compared to larger fragments.
Procedure:
1. Preparation of Gel: Agarose or polyacrylamide gel is commonly used for DNA electrophoresis. The gel is prepared by dissolving the appropriate amount of agarose or polyacrylamide in a buffer solution and pouring it into a gel tray with comb slots.
2. Loading the DNA Samples: The DNA fragments generated by the restriction endonucleases are mixed with a loading dye, which provides density and color to the sample. The mixture is loaded into the wells created by the comb in the gel.
3. Applying Electric Field: The gel tray is placed in an electrophoresis chamber filled with a buffer solution. The positive electrode is placed at one end, and the negative electrode is placed at the other end of the chamber. When the power supply is turned on, an electric field is established, and DNA fragments start migrating through the gel.
4. Visualization: To visualize the separated DNA fragments, the gel is stained with a dye such as ethidium bromide, which intercalates between the DNA bases and fluoresces under UV light. The DNA bands can be visualized and photographed using a UV transilluminator.
5. Analysis: The separated DNA fragments appear as distinct bands on the gel. The distance traveled by each band is measured from the origin. By comparing the migration distances of known DNA size markers, the size of the unknown DNA fragments can be estimated.
Advantages of Electrophoresis:
- It is a widely used technique for separating DNA fragments based on size.
- It is relatively simple and cost-effective.
- It can handle multiple samples simultaneously.
- It provides qualitative and semi-quantitative information about the size of DNA fragments.
Conclusion:
Electrophoresis is the most suitable method for separating DNA fragments generated by restriction endonucleases. It allows for the visualization and analysis of DNA fragments based on their size, enabling researchers to study genetic variations, perform DNA fingerprinting, and carry out various molecular biology techniques.