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Question
The lysate of cells scraped out of the buccal cavity was treated with all the macromolecule digesting enzymes except DNase. After purification, the resulting fraction of DNA was cut using enzymes. The technique used to separate the resulting fragments of DNA could be
I. gel electrophoresis
II. polymerase chain reaction
III. restriction digestion
IV. southern blotting
The lysate of cells scraped out of the buccal cavity was treated with all the macromolecule digesting enzymes except DNase. After purification, the resulting fraction of DNA was cut using enzymes. The technique used to separate the resulting fragments of DNA could be
I. gel electrophoresis
II. polymerase chain reaction
III. restriction digestion
IV. southern blotting
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Solution
The correct option is C only I
The extract containing all the components of the cell after breakdown of the cell wall and/or cell membrane using degradative enzymes is known as cell lysate.
The treatment of cells scraped out of the buccal cavity with macromolecule digesting enzymes except DNases (enzyme responsible for degradation of DNA molecules) leaves the DNA intact.
Cleavage of a DNA molecule yields several fragments, each containing a specific number of base pairs.Assuming that the fragments are of various sizes, those can be separated by a technique called gel electrophoresis.
Gel electrophoresis is a laboratory technique based on the principle of movement of charged particles towards oppositely charged electrodes under the influence of electric field. This is used for the separation of a mixture of DNA (a negatively charged molecule) fragments of varying sizes on agarose gel (a natural biopolymer) matrix under the influence of an electric field. The negatively charged DNA molecule moves from the negative terminal of the gel towards the positive terminal. The distance migrated by the DNA fragment depends on the size of the molecule. DNA with more base pairs can migrate to shorter distances than those with a lesser number of base pairs in a gel of a particular composition. Besides, the rate of migration is also determined by the pore size of the gel. The DNA fragments after separation form bands at different positions in the lane. These bands can be observed by using ethidium bromide as a stain and subsequently exposing the gel to ultraviolet light.
Polymerase chain reaction is a technique where a specific sequence of DNA can be amplified. It involves the use of pre-designed primers. These, then undergo cycles of denaturation, annealing and extension which gives an amplified amount of the parent DNA sequence. It is not used for separation of DNA fragments.
Restriction enzymes are those that cleave the DNA molecule at specific sequences after identifying them. These are used to generate fragments out of one DNA molecule. However, they do not have any role in separation of DNA.
Southern blotting is a technique which involves the transfer of DNA fragments from the agarose gel to nylon or nitrocellulose paper. This is then utilised to visualize or find out the desired DNA sequences by allowing hybridisation with a radioactive probe. This is commonly used in DNA fingerprinting technique.
The extract containing all the components of the cell after breakdown of the cell wall and/or cell membrane using degradative enzymes is known as cell lysate.
The treatment of cells scraped out of the buccal cavity with macromolecule digesting enzymes except DNases (enzyme responsible for degradation of DNA molecules) leaves the DNA intact.
Cleavage of a DNA molecule yields several fragments, each containing a specific number of base pairs.Assuming that the fragments are of various sizes, those can be separated by a technique called gel electrophoresis.
Gel electrophoresis is a laboratory technique based on the principle of movement of charged particles towards oppositely charged electrodes under the influence of electric field. This is used for the separation of a mixture of DNA (a negatively charged molecule) fragments of varying sizes on agarose gel (a natural biopolymer) matrix under the influence of an electric field. The negatively charged DNA molecule moves from the negative terminal of the gel towards the positive terminal. The distance migrated by the DNA fragment depends on the size of the molecule. DNA with more base pairs can migrate to shorter distances than those with a lesser number of base pairs in a gel of a particular composition. Besides, the rate of migration is also determined by the pore size of the gel. The DNA fragments after separation form bands at different positions in the lane. These bands can be observed by using ethidium bromide as a stain and subsequently exposing the gel to ultraviolet light.
Polymerase chain reaction is a technique where a specific sequence of DNA can be amplified. It involves the use of pre-designed primers. These, then undergo cycles of denaturation, annealing and extension which gives an amplified amount of the parent DNA sequence. It is not used for separation of DNA fragments.
Restriction enzymes are those that cleave the DNA molecule at specific sequences after identifying them. These are used to generate fragments out of one DNA molecule. However, they do not have any role in separation of DNA.
Southern blotting is a technique which involves the transfer of DNA fragments from the agarose gel to nylon or nitrocellulose paper. This is then utilised to visualize or find out the desired DNA sequences by allowing hybridisation with a radioactive probe. This is commonly used in DNA fingerprinting technique.
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