-EXERCISE-9

NAME: LOUIZA L.
ABINES BSED SCIENCES 3AX DATE: APRIL 26, 2023

INSTRUCTOR: PROF MERALUNA CANUNAYON
Student Web quest: PCR Source: Genetic Science Learning Center. "PCR Virtual Lab." Learn.Genetics
14
July 2009 http://learn.genetics.utah.edu/content/labs/pcr/
Objective: In this virtual lab, you will identify the resources and process of Polymerase Chain Reaction or
PCR.
Introduction:
1. How many base pairs are there in the human genome?

- The human genome consists of approximately 3.2 billion base pairs.
2. What is PCR? What does it do?
- PCR is a molecular biology technique that amplifies a specific segment of DNA using a DNA
template, primers, DNA polymerase, and nucleotides. It allows for the generation of millions or
billions of copies of the DNA segment of interest and is used in a wide range of biological
applications, including medical diagnostics, genetic research, and forensics.
3. What are some of the advantages of PCR? Name 1 and explain.
- Some advantages of PCR include its high specificity, sensitivity, and speed. One advantage of PCR
is its high specificity, which allows for the amplification of a specific DNA sequence while ignoring
another DNA present in the sample. This makes PCR a powerful tool for detecting pathogens or
genetic variations that may be present in very small amounts. Additionally, PCR can be performed
relatively quickly and requires only a small amount of DNA, making it a useful technique for many
applications in research and diagnostics.
4. What is unique about the design of PCR tubes?
- PCR tubes are designed with a special type of plastic that can withstand high temperatures required
for PCR reactions, a thin, flat wall for efficient heat transfer, and a tight-fitting cap to prevent
contamination and evaporation. They may also have a frosted surface for easy labeling and be
compatible with thermal cyclers.
At Lab Bench making the Mix: Note: Remember to click and hold the mouse to use the pipette.
5. Label the following on your bench. Your PCR tube (F) has already been done for you.
A DNA Extract D ______Nucleotides____
B &C _____Primer 1 and Primer 2____ E DNA Polymerase _
6. What is the first thing you need to add to your PCR tube? (Label A):
- To begin a PCR reaction, the first component that should be added to the PCR tube is the DNA
extract that contains the target DNA sequence to be amplified.
7. What is the function of a “primer?”

- The function of a primer in PCR is to provide a starting point for DNA synthesis by the DNA
polymerase enzyme. The primer binds to the complementary DNA sequence flanking the region of
interest and ensures that the DNA polymerase enzyme only amplifies the specific region of interest
during the PCR reaction.
8. What does DNA Polymerase do?

- DNA polymerase adds nucleotides to a growing DNA strand during DNA replication or PCR. It is
responsible for extending the primer and creating a new complementary strand of DNA during
PCR.
Inside of the Thermal cycler:
9.In Cycle 1 at 95 Degrees Celsius, what happens to the DNA?

- During Cycle 1 of PCR, the DNA is heated to 95 degrees Celsius,
which causes the double-stranded DNA to separate or denature into
two single-stranded pieces.
10.In Cycle 1 at 50 Degrees Celsius, what anneals (attaches) to the

DNA strands?
- During the annealing step of PCR, which typically occurs at
temperatures around 50-60 degrees Celsius, the primer binds or
attaches to the complementary single-stranded DNA template. This
allows the DNA polymerase enzyme to initiate DNA synthesis, which
leads to the amplification of the target DNA sequence.
11.In Cycle 1 at 72 Degrees Celsius, what is activated and what is

added to the DNA strands?
- During the extension phase of PCR, which typically occurs at around
72 degrees Celsius, the DNA polymerase enzyme is activated and
starts adding nucleotides to the 3' end of the primers on the
complementary single-stranded DNA template.
12.What are the end products of Cycles 2 and Cycles 3?

- After multiple cycles of denaturation, annealing, and extension in
PCR, the desired fragments of DNA are amplified. In particular, cycles
2 and 3 contribute to the exponential amplification of the target DNA
sequence, resulting in the production of a significant quantity of the
desired DNA fragment.
13.Label the processes of DNA replication shown to the left.
A. The primer is being attached to the single stranded DNA.

B. DNA polymerase attaches the complimentary strand
C. Nucleotides are added.
Analysis and Conclusions
14. Why do you think that everything on your lab bench had to be in ice?
- Everything on the lab bench had to be kept on ice to preserve the stability and activity of the reagents and
enzymes, prevent DNA contamination or degradation, and minimize temperature-related variability for
accurate and reproducible results.
15. Why did the pipette change tips every time? What is this important?
- The pipette tip was changed every time to prevent cross-contamination between samples and to ensure
accurate and precise measurements. This is important because even small amounts of contamination or
imprecision can significantly affect the PCR reaction, leading to inaccurate or unreliable results.

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NAME: LOUIZA L.

ABINES BSED SCIENCES 3AX DATE: APRIL 26, 2023

1. How many base pairs are there in the human genome?

2. What is PCR? What does it do?

3. What are some of the advantages of PCR? Name 1 and explain.

4. What is unique about the design of PCR tubes?

A DNA Extract D __Nucleotides

B &C _Primer 1 and Primer 2 E DNA Polymerase _

7. What is the function of a “primer?”

8. What does DNA Polymerase do?

9.In Cycle 1 at 95 Degrees Celsius, what happens to the DNA?

10.In Cycle 1 at 50 Degrees Celsius, what anneals (attaches) to the

11.In Cycle 1 at 72 Degrees Celsius, what is activated and what is

12.What are the end products of Cycles 2 and Cycles 3?

13.Label the processes of DNA replication shown to the left.

A. The primer is being attached to the single stranded DNA.

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