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Question
Arrange the given steps involved in recombinant DNA technology in the correct order.
A. Amplification of desired DNA sequence (gene of interest) with PCR modification
B. Ligation of gene of interest with the vector
C. Removal of RNA and proteins to purify DNA
D. Insertion of recombinant DNA into the host
E. Treatment of cell with cell wall digesting enzyme followed by disruption of the cell membrane
F. Digestion of DNA of interest and vector with suitable restriction enzyme
Arrange the given steps involved in recombinant DNA technology in the correct order.
A. Amplification of desired DNA sequence (gene of interest) with PCR modification
B. Ligation of gene of interest with the vector
C. Removal of RNA and proteins to purify DNA
D. Insertion of recombinant DNA into the host
E. Treatment of cell with cell wall digesting enzyme followed by disruption of the cell membrane
F. Digestion of DNA of interest and vector with suitable restriction enzyme
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Solution
The correct option is C E → C → A → F → B → D
Recombinant DNA is formed by combining genes of interest with carrier DNA molecules called vectors. The vectors act as vehicles for genes of interest which when introduced into the host cell can make its copies inside the host.
The steps of recombinant DNA technology are as follows:
1. Isolation of genetic material: Pure DNA is isolated from the cell as a result of the following treatments:-
a. Treatment with cell wall digesting enzyme: DNA is released from the cell by breaking open the cell wall. Bacterial cells can be digested by using lysozyme, plant cells by cellulase and fungal cells by chitinase.
b. Disruption of cell membrane: The cell membrane is disrupted by making use of chemical detergents such as sodium dodecyl sulphate (SDS).
c. Treatment with RNases and proteases: The released DNA will be associated with other macromolecules like RNA and proteins. Ribonucleases (RNAases) and proteases can be used to digest them respectively to purify DNA.
2. Polymerase chain reaction (PCR): This is done in order to amplify the DNA fragment of interest as well as modify their ends.
3. Gel electrophoresis: It is a technique used to separate macromolecules based on their size and charge. This is done for the separation of DNA molecules of varying sizes. The portion of the gel that contains our DNA of interest is excised. The DNA is then extracted from the excised piece of gel by a process known as elution followed by their purification.
4. Restriction enzyme digestion of DNA of interest and the vector DNA: Common restriction enzymes are used to make cuts at specific sites on DNA to generate complementary sticky ends.
5. Ligation of gene of interest with vector: It is the joining of the two DNA fragments - gene of interest and the vector together with the help of the enzyme DNA ligase.
6. Insertion of recombinant DNA into host: In this step, the recombinant DNA is introduced into a host cell where it gets multiplied and is expressed in the form of protein under optimal conditions. This can be used for commercial purposes.
Recombinant DNA is formed by combining genes of interest with carrier DNA molecules called vectors. The vectors act as vehicles for genes of interest which when introduced into the host cell can make its copies inside the host.
The steps of recombinant DNA technology are as follows:
1. Isolation of genetic material: Pure DNA is isolated from the cell as a result of the following treatments:-
a. Treatment with cell wall digesting enzyme: DNA is released from the cell by breaking open the cell wall. Bacterial cells can be digested by using lysozyme, plant cells by cellulase and fungal cells by chitinase.
b. Disruption of cell membrane: The cell membrane is disrupted by making use of chemical detergents such as sodium dodecyl sulphate (SDS).
c. Treatment with RNases and proteases: The released DNA will be associated with other macromolecules like RNA and proteins. Ribonucleases (RNAases) and proteases can be used to digest them respectively to purify DNA.
2. Polymerase chain reaction (PCR): This is done in order to amplify the DNA fragment of interest as well as modify their ends.
3. Gel electrophoresis: It is a technique used to separate macromolecules based on their size and charge. This is done for the separation of DNA molecules of varying sizes. The portion of the gel that contains our DNA of interest is excised. The DNA is then extracted from the excised piece of gel by a process known as elution followed by their purification.
4. Restriction enzyme digestion of DNA of interest and the vector DNA: Common restriction enzymes are used to make cuts at specific sites on DNA to generate complementary sticky ends.
5. Ligation of gene of interest with vector: It is the joining of the two DNA fragments - gene of interest and the vector together with the help of the enzyme DNA ligase.
6. Insertion of recombinant DNA into host: In this step, the recombinant DNA is introduced into a host cell where it gets multiplied and is expressed in the form of protein under optimal conditions. This can be used for commercial purposes.
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