iGEM Thessaloniki - PCR

What of the following is not a required ingredient for PCR?

• Primers

• A

• DNA polymerase

• U

Which of the following is the correct order of thermal cycles?

• 94-72-50

• 94-50-72

• 72-50-94

• 72-94-50

After four cycles how many copies will have been produced (considering that we have n copies when we start)?

• 4n

• 8n

• 16n

• 32n

Which of the following is not applicable for use in various PCR techniques?

• dNTPs

• Polymerase from human body cells

• RNA strand

• Primer of 20b length

At what temperature does the Denature step happen?

• 94

• 72

• 50

• 24

What is the function of a primer?

• To identify the particular region of DNA to be copied by PCR.

• To maintain the temperature of the PCR reaction.

• To create DNA nucleotides.

• To copy DNA.

What is the function of DNA Polymerase?

• Attach primers to target sequence

• Separate the DNA

• Attach nucleotides to the target sequence

• Separate the fragments according to size

PCR happens in 3 steps (denaturing, annealing, extending) or 1 "cycle". How many times are cycles typically repeated?

• Over 1 billion

• 1

• 30

• 10

What are some adjustable parameteres in a PCR experiment?

• Thermal cycler capabilities

• Annealing temperature

• Primer concentration

• MgCl2 concentration

When designing primers we must consider :

• GC%

• Tm of both primers (up/dowm)

• AT%

• All the above

Answer explanation

Consider the primers' stability and hybrization!

Primers must have similar Tm so that w can properly adjust the temperature (Annealing Step)

Primers must contain less than 55% GC. Their annealing mustn't be too strong.