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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
Answer the following question:
Describe the formation of recombinant DNA by the action of EcoRI.
In a pathological lab, a series of steps were undertaken for finding the gene of interest. Describe the steps, or make a flow chart showing the process of amplification of this gene of interest.
PCR and restriction fragment length polymorphism are the methods for ______.
PCR proceeds mthree distinct steps governed by temperature, they are in order of ______.
Significance of 'heat shock' method in bacterial transformation is to facilitate ______.
Which of the following should be chosen for best yield if one were to produce a recombinant protein in large amounts?
Read the following paragraph and answer the questions that follow:
| Biotechnology revolves around the "gene of interest", with an objective to open various avenues for human welfare in health, medicine, pharma, agriculture etc. using different techniques, tools and processes. One of the breakthroughs of biotechnology in medicine is the gene therapy. |
- Name the human disease for which the gene therapy was used for the first time.
- Explain the steps of gene therapy carried to cure the disease using the lymphocytes of the patient. Why is this therapy not a permanent cure of the disease?
- Write the possible permanent cure of the disease by the gene therapy that is in progress.
Upon exposure to UV radiation, DNA stained with ethidium bromide will show ______.
Which enzyme is utilized in Polymerase Chain Reaction (PCR) because it remains active at high temperatures?
How are bacterial cells typically made competent to facilitate the uptake of recombinant DNA during transformation?
