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प्रश्न
Explain briefly:
PCR
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उत्तर
PCR (Polymerase Chain Reaction) is a technique used to create multiple copies of a specific DNA segment in vitro. It amplifies DNA by repeated cycles of three steps:
- Denaturation: Heating the double-stranded DNA to around 94°C to separate it into single strands.
- Annealing: Cooling to 46-60°C to allow short primers to bind to complementary sequences on the single-stranded DNA.
- Extension: Raising the temperature to about 72°C so a heat-stable DNA polymerase (Taq polymerase) synthesizes new complementary strands, duplicating the target DNA.
Repeating these cycles exponentially amplifies the DNA segment, producing millions of copies rapidly. PCR is widely used for DNA amplification, genetic fingerprinting, disease detection, DNA sequencing, and molecular mapping. It was introduced by Kary Mullis in 1985 and relies on the thermostable enzyme from Thermus aquaticus to function at high temperatures without being denatured.
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संबंधित प्रश्न
Suggest and describe a technique to obtain multiple copies of a gene of interest in vitro.
Draw a labelled sketch of sparged-stirred-tank bioreactor. Write its application.
Explain briefly:
Chitinase
State the importance of using a bioreactor.
Prepare a flow chart in formation of recombinant DNA by the action of restriction endonuclease enzyme EcoRI.
Answer the following question:
Describe the formation of recombinant DNA by the action of EcoRI.
DNA fragments generated by the restriction endonuclease in a chemical reaction can be separated by ______.
PCR and restriction fragment length polymorphism are the methods for ______.
PCR proceeds mthree distinct steps governed by temperature, they are in order of ______.
The first step in recombinant DNA technology is
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?
Rising of dough is due to ______.
How is copy number of the plasmid vector related to yield of recombinant protein?
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.
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.
State the advantage of using Thermostable DNA polymerase.
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.
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:
In agarose gel electrophoresis, why do DNA fragments move toward the positive electrode?
Which enzyme is responsible for joining the desired gene to a carrier vector DNA to form recombinant DNA?
How are bacterial cells typically made competent to facilitate the uptake of recombinant DNA during transformation?
