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प्रश्न
Suggest and describe a technique to obtain multiple copies of a gene of interest in vitro.
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उत्तर
For the PCR, two sets of primers (chemically synthesised oligonucleotides that are complementary to a region of DNA), the enzyme DNA polymerase and deoxynucleotides are added.
The PCR consists of three steps:
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Denaturation: Double helical DNA is denatured at a high temperature. DNA polymerase does not get degraded at this temperature, as the DNA polymerase used in this reaction is thermostable. It is isolated from the thermophilic bacterium Thermus aquaticus (Taq).
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Annealing: It is the step in which primers are annealed to single-stranded DNA templates. Two sets of primers (chemically synthesised small oligonucleotides that are
complementary to the regions of DNA) are used. The temperature of the reaction mixture is lowered to 50–65 °C for some seconds to allow annealing of the primers. DNA polymerase extends the primer in the 5' to 3' direction. -
Extension: Replication of DNA occurs in vitro.
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This cycle is repeated several times to generate up to one billion identical copies of DNA.
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संबंधित प्रश्न
Name two commonly used bioreactors.
Can you recall meiosis and indicate at what stage recombinant DNA is made?
Explain briefly:
Chitinase
______ plays an important role in blood clotting.
DNA fragments generated by the restriction endonuclease in a chemical reaction can be separated by ______.
Gene Amplification using primers can be done by ______.
PCR proceeds mthree distinct steps governed by temperature, they are in order of ______.
How is copy number of the plasmid vector related to yield of recombinant protein?
Main steps in the formation of Recombinant DNA are given below. Arrange these steps in a correct sequence.
- Insertion of recombinant DNA into the host cell.
- Cutting of DNA at specific location by restriction enzyme.
- Isolation of desired DNA fragment.
- Amplification of gene of interest using PCR.
Choose the correct answer from the options given below:
Upon exposure to UV radiation, DNA stained with ethidium bromide will show ______.
