Polymerase Chain Reaction

What needs to be done to the sample of blood before PCR?

DNA must be isolated from the cells.

How did you collect quids in the lab?

A pipette

What do you need to do each time before using a pipette to collect liquid?

Put on a new, sterile pipette tip.

What is the function of primers in a PCR reaction?

Bind specific sites on DNA

Great that you throw away the used pipette tip, why is it important to change the pipette tip?

to avoid cross contamination

For which enzymes are nucleotides the substrate?

DNA Polymerase

What is the template of the PCR reaction?

DNA

What does DNA polymerase do?

Synthesizes DNA

AT this step in the DNA process, what happens to the DNA?

It will be separated into two stands.

How is the DNA separated into a single strand?

The high temperature 95° separates them.

What is the step in the PCR reaction that is now shown called?

Annealing

The area where the primers bind marks which part of the PCR product?

Beginning

The PCR products get a certain length due to which fact?

The placement of the primers

How does the DNA polymerase extend the primers into a new DNA strand?

A Nucleotides to the 3' end of the primers

Which of these sequences is the complimentary sequence to the template sequence:5'-GTGGTCTGATCAACGGTAA-3'

c)

3'-CACCAGACTAGTTGCCATT-5'

When using one primer pair in different individuals, which phrase describes the PCR found?

Different length may be found in each individual

What happens to the probability of a 100% match between two different individuals when using 13 sets of primers for the DNA profile instead of one?

It decreases

How many copies of DNA are required to see bands on the electrophoresis gel?

Millions of copies

Which word describes the charge of the DNA?

Negatively charged

DNA is negatively charged. To which location in the electrophoresis gel does it migrate?

The Positive pole

So, who was the murderer?

The rival scientist

(The murder strand matches the rival scientist strand)

What are the building blocks of new copies of DNA?

Nucleotides

What is the function of primers in a PCR?

They bind specific sites on the template DNA to initiate and direct DNA synthesis

What would happen if no polymerase was added to the PCR reaction?

New DNA would not be generated

Which reagant acts as a template for the DNA polmerase so that it knows which new DNA to make?

DNA from the blood sample

DNA polymerase bonds to the template DNA. In which direction is the new DNA subsequently synthesized?

5'-3'

Why is the Taq-polymerase special compared to most other polymerases?

It can resist high temperatures

What can contamination of reagents lead to?

Unreliable results

How many sets of primers are needed for DNA profiling?

13

What is the purpose of PCR?

To copy and then make many copies of a specific region of DNA

Why is a PCR cycle repeated 30 times?

To get enough DNA

What can a DNA ladder help determine?

The length of a fragment

Why is it possible to distinguish different individuals by running these PCR products on a gel?

The PCR products are different lengths