16 Questions
What is the minimum number of base pairs required for restriction endonucleases to function?
Over 1000
Which type of restriction endonucleases are most important in gene cloning?
Type II
What does the enzyme DNase I target in a DNA molecule?
Any internal phosphodiester bond
Which enzyme attaches to a short single-stranded region in a mainly double-stranded DNA molecule?
DNA polymerase I
What are sticky ends in DNA manipulation?
Ends of DNA that can form base pairs with complementary sequences
What is the function of the Klenow fragment?
To synthesize new DNA strands without degrading existing ones
Which enzyme is thermostable and used in PCR?
Taq DNA polymerase
What are linkers in DNA manipulation?
Short pieces of double-stranded DNA with a known sequence
What is the primary purpose of gene cloning experiments?
To construct recombinant DNA molecules
What is the function of nucleases in DNA manipulation?
To degrade DNA molecules by breaking phosphodiester bonds
Which enzyme is used to synthesize a DNA strand complementary to an RNA template?
Reverse transcriptase
What type of enzyme is used to remove phosphate groups from the 5′ terminus of a DNA molecule?
Alkaline phosphatase
Which type of nuclease removes nucleotides one at a time from the end of a DNA molecule?
Exonucleases
What is the role of DNA ligase during DNA replication?
To repair single-stranded breaks in DNA
Which enzyme is commonly used in the polymerase chain reaction (PCR)?
Taq DNA polymerase
What are the two examples of DNA manipulative techniques mentioned?
Cutting and joining DNA
Study Notes
Gene Cloning Experiments
- The primary purpose of gene cloning experiments is to construct recombinant DNA molecules.
- Gene cloning involves DNA manipulation techniques, such as cutting and joining DNA.
DNA Manipulation Enzymes
- Nucleases are enzymes that degrade DNA molecules by breaking phosphodiester bonds.
- Ligase enzymes are involved in joining nucleic acid molecules together.
- DNA polymerase is an enzyme used in the polymerase chain reaction (PCR).
- Alkaline phosphatase is an enzyme that removes phosphate groups from the 5′ terminus of a DNA molecule.
- Exonucleases are enzymes that remove nucleotides one at a time from the end of a DNA molecule.
- Endonucleases are enzymes that cleave DNA at specific internal sites.
Specific Enzymes
- Taq DNA polymerase is a thermostable enzyme commonly used in PCR.
- Reverse transcriptase is an enzyme that synthesizes a DNA strand complementary to an RNA template.
- Terminal deoxynucleotidyl transferase is an enzyme that adds deoxyribonucleotides onto the 3′ terminus of a DNA molecule.
- S1 endonuclease is an enzyme purified from the fungus Aspergillus oryzae.
- Deoxyribonuclease I (DNase I) is an enzyme that targets any internal phosphodiester bond in a DNA molecule.
DNA Restriction and Ligation
- Over 1000 different restriction endonucleases have been isolated.
- Type II restriction endonucleases are the most important in gene cloning.
- Sticky ends are ends of DNA that can form base pairs with complementary sequences.
- Linkers are short pieces of double-stranded DNA with a known sequence, used in DNA manipulation.
- A problem that can arise when using linkers is that they can form dimers.
This quiz tests your understanding of gene cloning, DNA manipulation, and enzymes involved in DNA processes.
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