Target Exam

CUET

Subject

Biology

Chapter

Biotechnology -Principles and Processes

Question:

Read the following paragraph and answer the given questions.

Repeated amplification is achieved by the use of a thermostable DNA polymerase (isolated from a bacterium, Thermus aquaticus), which remain active during the high temperature induced denaturation of double stranded DNA. The separated DNA fragments can be visualised only after staining the DNA with a compound known as ethidium bromide followed by exposure to UV radiation. The separated bands of DNA are cut out from the agarose gel and extracted from the gel piece. The convention for naming these enzymes is the first letter of the name comes from the genus and the second two letters come from the species of the prokaryotic cell from which they were isolated, e.g., EcoRI comes from Escherichia coli RY 13. In EcoRI, the letter 'R' is derived from the name of strain. Roman numbers following the names indicate the order in which the enzymes were isolated from that strain of bacteria. Stanley Cohen and Herbert Boyer accomplished this in 1972 by isolating the antibiotic resistance gene by cutting out a piece of DNA from a plasmid which was responsible for conferring antibiotic resistance. Since the DNA is enclosed within the membranes, we have to break the cell open to release DNA, RNA, protein, polysaccharides etc. The RNA can be removed by treatment with ribonuclease whereas proteins can be removed by treatment with protease. Other molecules can be removed by appropriate treatments and purified DNA ultimately precipitates out after the addition of chilled ethanol.

In the isolation process of genetic material, the purified DNA can be precipitated out after the addition of:

Options:

Chilled water

Chilled methanol

Chilled ethanol

Warm ethanol

Correct Answer:

Chilled ethanol

Explanation:

The correct answer is Option (3) -Chilled ethanol

Nucleic acid is the genetic material of all organisms without exception. In majority of organisms this is deoxyribonucleic acid or DNA. In order to cut the DNA with restriction enzymes, it needs to be in pure form, free from other macro-molecules.Since the DNA is enclosed within the membranes, we have to break the cell open to release DNA along with other macromolecules such as RNA, proteins, polysaccharides and also lipids. This can be achieved by treating the bacterial cells/plant or animal tissue with enzymes such as lysozyme (bacteria), cellulase (plant cells), chitinase (fungus). Genes are located on long molecules of DNA interwined with proteins such as histones. The RNA can be removed by treatment with ribonuclease whereas proteins can be removed by treatment with protease. Other molecules can be removed by appropriate treatments and purified DNA ultimately precipitates out after the addition of chilled ethanol. This can be seen as collection of fine threads in the suspension.Isolation of purified DNA involves the use of chilled ethanol to precipitate DNA out of a mixture of biomolecules, allowing for the separation and purification of DNA.