MIDTERM MOLBIO LEC GENE EXPRESSION

The process of turning on a gene to produce RNA and protein.

• DNA Replication

• Gene Expression

• Gene Regulation

• Transcription

• Translation

Arrange the following PCR steps:

I. Initialization

II. Annealing

III. Extension/Elongation

IV. Denaturation

• I - IV - II - III

• I - II - III - IV

• I - III - II - IV

• I - IV - III - II

This PCR step is only required for DNA polymerases that require heat activation by hot-start PCR.

• Initialization

• Denaturation

• Annealing

• Extension/Elongation

This step is the first regular cycling event and consists of heating the reaction chamber which causes DNA melting, or denaturation, of the double-stranded DNA template by breaking the hydrogen bonds between complementary bases, yielding two single-stranded DNA molecules.

• Initialization

• Denaturation

• Annealing

• Extension/Elongation

The polymerase binds to the primer-template hybrid and begins DNA formation.

• Initialization

• Denaturation

• Annealing

• Extension/Elongation

The DNA polymerase synthesizes a new DNA strand complementary to the DNA template strand by adding free dNTPs from the reaction.

• Initialization

• Denaturation

• Annealing

• Extension/Elongation

During initialization step in PCR, the reaction chamber is heated to a temperature of ____________ if extremely thermostable polymerases are used, which is then held for ____________

• 94-96 °C or 98; 1–10 minutes

• 94– 98 °C; 20- 30 seconds

• 50–65 °C; 20–40 seconds

• 75–80 °C

During denaturation step in PCR, the reaction chamber is heated to a temperature of ____________ for ____________ which causes DNA melting, or denaturation, of the double-stranded DNA template

• 94-96 °C or 98; 1–10 minutes

• 94– 98 °C; 20- 30 seconds

• 50–65 °C; 20–40 seconds

• 75–80 °C

During annealing step in PCR, the reaction temperature is lowered to _____________ for ________________, allowing annealing of the primers to each of the single-stranded DNA templates

• 94-96 °C or 98; 1–10 minutes

• 94– 98 °C; 20- 30 seconds

• 50–65 °C; 20–40 seconds

• 75–80 °C

During extension/elongation step in PCR, the optimum activity temperature for the thermostable DNA polymerase of Taq polymerase is approximately

• 94-96 °C or 98; 1–10 minutes

• 94– 98 °C; 20- 30 seconds

• 50–65 °C; 20–40 seconds

• 75–80 °C