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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
Describe briefly the following:
Bioreactors
State the importance of using a bioreactor.
Prepare a flow chart in formation of recombinant DNA by the action of restriction endonuclease enzyme EcoRI.
Gene Amplification using primers can be done by ______.
PCR proceeds mthree distinct steps governed by temperature, they are in order of ______.
The first step in recombinant DNA technology is
While doing a PCR, ‘denaturation’ step is missed. What will be its effect on the process?
Illustrate the design of a bioreactor. Highlight the difference between a flask in your laboratory and a bioreactor which allows cells to grow in a continuous culture system.
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.
Bioreactors are the containment vehicles of any biotechnology-based production process. For large scale production and for economic reasons the final success of biotechnological process depends on the efficiency of the bioreactor.
Answer the following questions w.r.t. the given paragraph:
- List the operational guidelines that must be adhered to so as to achieve optimisation of the bioreactor system. Enlist any four.
- Mention the phase of the growth we refer to in the statement "Optimisation of growth and metabolic activity of the cells".
- Is the biological product formed in the bioreactor suitable for the intended use immediate? Give reason in support of your answer.
