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Questions
Distinguish between exonuclease and endonuclease.
Differentiate between exonuclease and endonuclease.
Discuss with your teacher and find out how to distinguish between exonuclease and endonuclease.
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Solution 1
| Sr. No. | Exonuclease | Endonuclease |
| 1. | It is a type of restriction enzyme that removes the nucleotide from the 5' or 3' ends of the DNA molecule. | It is a type of restriction enzyme that makes a cut within the DNA at a specific site to generate sticky ends. |
| 2. | They separated DNA base pairs at their terminal ends. |
They separate DNA at any location other than the terminal ends. |
| 3. | They work on single strands of DNA or gaps in double-stranded DNA. | They cut one or both strands of double-stranded DNA. |
Solution 2
| Feature | Exonuclease | Endonuclease |
| Clevage Site: | Eliminates nucleotides from DNA/RNA ends. | Cuts DNA within the strand at specific sites. |
| Requirement of Free Ends: | Needs free 3' or 5' ends (in one duplex strand). | Requires just one or both strands of the DNA duplex and doesn’t require free ends. |
| Mode of Action: | Progressively removes nucleotides one by one. | Cuts at specific recognition sequences. |
| Example: | DNA polymerase I (has exonuclease activity). | EcoRI, HindIII, BamHI (restriction enzymes). |
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(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
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- smallest/lightest DNA strand in the matrix
- Agarose gel
State the importance of elution in this process.
Given below is the restriction site of a restriction endonuclease Pst-I and the cleavage sites on a DNA molecule.
\[\ce{5' C - T - G - C - A \overset{\downarrow}{-}{G 3'}}\]
\[\ce{3' G\underset{\uparrow}{-} A - C - G - T - C 5'}\]
Choose the option that gives the correct resultant fragments by the action of the enzyme Pst-I.
'EcoRI' has played a very significant role in rDNA technology.
- Explain the convention for naming EcoRI.
- Write the recognition site and the cleavage sites of this restriction endonuclease.
What are the protruding and hanging stretches of DNA produced by these restriction enzymes called? Describe their role in the formation of rDNA.
