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प्रश्न
Name and describe the technique that helps in separating the DNA fragments formed by the use of restriction endonuclease
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उत्तर
Gel electrophoresis is a technique of separating DNA fragments formed by the action of restriction endonucleases.
The fragments of DNA are placed in a typical agarose gel under an electric field. The DNA fragments move towards the anode as these fragments are negatively charged molecules. The DNA fragments separate according to their size through the sieving effect provided by the agarose gel. The smaller the fragment size, the farther it moves. The separated DNA fragments are stained with ethidium bromide followed by exposure to ultraviolet radiation. The DNA fragments are seen as orangecoloured and are cut out from the agarose gel and extracted from the gel piece. This step is called elution.
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संबंधित प्रश्न
Mention the difference in the mode of action of exonuclease and endonuclease.
Collect 5 examples of palindromic DNA sequences. Better try to create a palindromic sequence by following base-pair rules.
The total number of nucleotide sequences of DNA that code for a hormone is 1530. The proportion of different bases in the sequence is found to be Adenine = 34%, Guanine = 19%, Cytosine = 23%, Thymine = 19%.
Applying Chargaff’s rule, what conclusion can be drawn?
'Restriction' in restriction enzyme refers to
DNA strands on a gel stained with ethidium bromide when viewed under UV radiation, appear as ______
Restriction enzymes should not have more than one site of action in the cloning site of a vector. Comment.
Carefully observe the given picture. A mixture of DNA with fragments ranging from 200 base pairs to 2500 base pairs was electrophoresed on agarose gel with the following arrangement.
(a) What result will be obtained on staining with ethidium bromide? Explain with reason.
(b) The above setup was modified and a band with 250 base pairs was obtained at X.
What change(s) were made to the previous design to obtain a band at X? Why did the band appear at position X?
Given below is the stepwise schematic representation of the process of electrophoresis. Identify the 'alphabets' representing
- Anode end
- smallest/lightest DNA strand in the matrix
- Agarose gel
State the importance of elution in this process.
What are the protruding and hanging stretches of DNA produced by these restriction enzymes called? Describe their role in the formation of rDNA.
