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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
Name two commonly used bioreactors.
Besides better aeration and mixing properties, what other advantages do stirred tank bioreactors have over shake flasks?
Can you recall meiosis and indicate at what stage recombinant DNA is made?
Describe briefly the following:
Downstream processing
Explain briefly:
Chitinase
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.
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.
DNA fragments generated by the restriction endonuclease in a chemical reaction can be separated by ______.
A single strand of nucleic acid tagged with a radioactive molecule is called ______.
PCR proceeds mthree distinct steps governed by temperature, they are in order of ______.
Rising of dough is due to ______.
Which of the following steps are catalysed by Taq DNA polymerase in a PCR reaction?
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 ______.
Which of the following should be chosen for best yield if one were to produce a recombinant protein in large amounts?
How is copy number of the plasmid vector related to yield of recombinant protein?
Identify and explain steps ‘A’, ‘B’ and ‘C’ in the PCR diagram given below.
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.
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.
Given below are two statements:
Statement I: The DNA fragments extracted from gel electrophoresis can be used in construction of recombinant DNA.
Statement II: Smaller size DNA fragments are observed near anode while larger fragments are found near the wells in an agarose gel.
In the light of the above statements, choose the most appropriate answer from the options given below:
What is the process of cutting out and recovering a separated DNA band from an agarose gel called?
In the provided agarose gel electrophoresis image, what is represented in lane 1 compared to lanes 2 through 4?
Which enzyme is utilized in Polymerase Chain Reaction (PCR) because it remains active at high temperatures?
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?
