Replication and manipulation

Question 1

Telomerase

Answer 1

Enzyme containing RNA complementary to the telomere sequence.
- Replaces the lost telomere repeats by adding DNA nucleotides to the 3’ end of the template strand.

Question 2

Okazaki fragments

Answer 2

Short pieces in the lagging strand.

Question 3

Polymerase proofreading

Answer 3

Detects bases that are misfired, “checks work” before DNA polymerase adds a new base.

Question 4

Polymerase chain reaction (PCR)

Answer 4

Used to amplify the DNA sample.

- Allows a single or a few copies of DNA to be replicated into millions or billions of copies.

Question 5

Restriction endonuclease (restriction enzymes)

Answer 5

Nucleotide sequence that cuts the DNA at a specific site.

Question 6

Gel electrophoresis

Answer 6

Smaller fragments move faster and larger fragments move slower.

Question 7

Southern blot
(DNA fingerprinting)

Answer 7

• A restriction enzyme is added to a solution containing the DNA.
• Fragments are separated by size through gel electrophoresis.
• DNA fragments in the gel are transferred (blotted) onto filter paper.
• Filter paper is removed and placed into a plastic bag containing a solution.
• Probe sticks to complementary fragments.
• Filter paper is exposed to X-ray film to darken the film over each band and help visualize their positions.

Question 8

Northern blot

RNA

Answer 8

Same idea but with RNA.

Question 9

Western blot

proteins

Answer 9

Same idea but with proteins.

Question 10

Sanger sequencing

Answer 10

Helps determine the DNA sequence.

• Dideoxynucleotides are added to a tube of a DNA sequence.
• Differing strands of DNA are formed and then separated through gel electrophoresis.
• DNA sequence can be determined, can use fluorescent dye.

Question 11

Bacterial transformation

Answer 11

Recombinant DNA where the offhanging strands are joined.

- Circular and linear.

Question 12

DNA polymerase

Answer 12

Enzyme responsible for forming new copies of DNA.