Question

Restriction enzymes recognize specific DNA sequences and cut each strand of DNA at specific locations at the target sequence. The result of digesting a particular genome with a particular restriction enzyme is a collection of restriction fragments of defined length and composition. These can be used to generate restriction maps or create pieces with sticky ends. These sticky ends can be used to attach to other fragments that have sticky ends caused by cutting with a different restriction enzyme. These pieces can be joined permanently by using ligase to create recombinant pieces for cloning. Selected Coordinates Clear
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Answer 1

1. Restriction enzymes recognize specific DNA sequences and cut each strand of DNA at specific locations at the target sequence.- Restriction enzyme cuts double stranded DNA at specific nucleotide sequence on target sequence. Single cut on each strand at specific location on target sequence

2. The result of digesting a particular genome with a particular restriction enzyme is a collection of restriction fragments of defined length and composition. - Digestion will result in fragments of defined length provided host controlled modification of bases like methylation of bases at target sequence is absent . Also digestion is sequence specific and changes in sequences other than recognition and target sequence, due to mutation may alter composition.

3. These can be used to generate restriction maps or create pieces with sticky ends. Correct. Restriction enzymes creates sticky ends as well as blunt ends depending on enzyme used.

4. These sticky ends can be used to attach to other fragments that have sticky ends caused by cutting with a different restriction enzyme. Incorrect, since restriction enzymes are site specific, use of different restriction enzyme may have different recognition and target sequence. Thus, sticky ends caused by same restriction enzyme with complementary bases on it will be essential for ligation

5. These pieces can be joined permanently by using ligase to create recombinant pieces for cloning. DNA ligase enzyme act on 5' phosphate groups of DNA molecules (with complementary nucleotide base pairing between two pieces of DNA) to form phosphodiester bond between two DNA molecules to generate recombinant DNA molecule.

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