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प्रश्न
Name and explain the technique used for separating DNA fragments and making them available for biotechnology experiments.
Name and describe the technique that helps in the separation and isolation of DNA fragments.
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उत्तर
Gel electrophoresis is a technique which is used for separating DNA fragments in a gel on the basis of their electric charge and size.
The process of gel electrophoresis is as follows:
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The fragments obtained after cutting with restriction enzymes are separated by using gel electrophoresis.
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Electric field is applied to the agarose gel matrix and the negatively-charged DNA fragments move towards the anode.
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Fragments separate according to their size by the sieving properties of agarose gel. The smaller the fragment, the farther it moves.
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Staining dyes such as ethidium bromide, upon exposure to UV radiation, are used to visualise the DNA fragments.
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DNA fragments are visible as bright orange-coloured bands in the agarose matrix.
- These bands are cut from the agarose gel and extracted from the gel piece (elution).
- DNA fragments are purified and used in constructing recombinant DNAs
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संबंधित प्रश्न
Give the steps in PCR or polymerase chain reaction with suitable diagrams.
Enlist different types of restriction enzymes commonly used in rDNA technology? Write about their role.
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Which of the following is an INCORRECT match?
Polymerase Chain Reaction is made possible by ____________.
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Who among the following was awarded the Nobel Prize for the development of PCR technique?
What would happen when one grows a recombinant bacterium in a bioreactor but forget to add antibiotic to the medium in which the recombinant is growing?
Cloning of genes, play a very significant role in genetic engineering, helping the transfer of desirable foreign genes into different hosts. The scientists, to make this process easier and effective are creating engineered vectors in such a way that they help easy linking of foreign DNA and selection of recombinants from non-recombinants. 'pBR322' is one such engineered vectors developed by scientists. A diagram of an engineered vector pBR322 is given below:
- Name the host for this cloning vector.
- Identify 'Rop' and 'Ori' in the diagram from 'U', 'V', 'W', 'X', 'Y' and 'Z'. Write their functions.
- Draw the fragments that will be formed by the action of 'Z' (marked in the diagram) on the specific site of the DNA segment given below:
5' --- GTACGAATCCTGA --- 3'
3' --- CATGCTTAGGACT --- 3'
Observe the given sequence of nitrogenous bases on a DNA fragments and answer the following questions:
5' - CAGAATTCTTA - 3'
3' - GTCTTAAGAAT - 5'
- Name a restriction enzyme which can recognise this DNA sequence.
- Write the sequence after digestion.
- Why are the ends generated after digestion called sticky ends?
Identify the desirable characteristics for a plasmid used in rDNA technology from the following.
- Ability to multiply and express outside the host in a bioreactor.
- A highly active promoter.
- A site at which replication can be initiated.
- One or more identifiable marker genes.
- One or more unique restriction sites.
Which of these are most widely used in genetic engineering?
To be a cloning vector, a plasmid require ______.
The polymerase chain reaction is a powerful technique to ______.
How gene amplification is done?
Write steps involved in gene amplification.
State the steps involved in the process of gene therapy for the treatment of ADA-deficiency.
Which of the following best describes the relationship between genetic engineering and recombinant DNA?
