Read the following paragraph carefully and answer the given questions. In the year 1963 the two enzymes responsible for restricting the growth of bacteriophage in E. coli were isolated. The first restriction endonuclease - Hind II, whose functioning depended on a specific DNA nucleotide sequence was characterized five years later. It was found that Hind II always cut DNA molecules at a particular point by recognizing a specific sequence of six base pairs. This specific base sequence is known as the recognition sequence for Hind II. Besides Hind II, today we know more than 900 restriction enzymes that have been isolated from over 230 strains of bacteria, each of which recognize different recognition sequences. |
DNA fragments can be separated by a technique known as: |
Autoradiography ELISA Ultracentrifugation Gel electrophoresis |
Gel electrophoresis |
The correct answer is Option (4) → Gel electrophoresis The process of cutting DNA using restriction endonucleases results in the formation of DNA fragments. These fragments can be separated using a technique called gel electrophoresis. Since DNA fragments are negatively charged molecules, they can be separated by applying an electric field, causing them to move towards the anode through a medium or matrix. Agarose, a natural polymer extracted from seaweeds, is commonly used as the matrix. The DNA fragments separate based on their size due to the sieving effect provided by the agarose gel. Smaller fragments move farther compared to larger fragments. The separated DNA fragments can be visualized only after staining the DNA with a compound called ethidium bromide, followed by exposure to UV radiation. Without staining, the pure DNA fragments cannot be seen in visible light. After staining, you can observe bright orange-colored bands of DNA in an ethidium bromide stained gel exposed to UV light. Once the DNA fragments are separated, they can be cut out from the agarose gel and extracted from the gel piece. This step is known as elution. The purified DNA fragments obtained through elution are then used in constructing recombinant DNA by joining them with cloning vectors.
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