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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.
Describe the process of amplification of the “gene of interest” using the PCR technique.
Which of the following term used for defining a viral genome incorporated into host DNA?
The first step in recombinant DNA technology is
In PCR, primers are used for ______.
During the process of gene amplification using PCR, if a very high temperature is not maintained in the beginning, then which of the following steps of PCR will be affected first?
Significance of 'heat shock' method in bacterial transformation is to facilitate ______.
A bacterial cell was transformed with a recombinant DNA molecule that was generated using a human gene. However, the transformed cells did not produce the desired protein. Reasons could be ______.
Assertion (A): Synthetic oligonucleotide polymers are used during Annealing in a PCR.
Reason (R): The primers bind to the double stranded DNA at their complementary regions.
Which of the following statements is incorrect?
