Answer key

DNA Replication Lab Instructions

DNA Replication: Data & Observations

Part I: Replicating DNA

Launch Bioscope
In Part I, you will watch as the two strands of the original DNA molecule separate and serve as
templates for the synthesis of new strands. You will also investigate the need for adenine (A),
thymine (T), guanine (G) and cytosine (C), from which DNA is made.

A. Click on Replicating DNA above the DNA viewer.

B. Examine the DNA molecule in the DNA Viewer and answer the following questions. Use
the Zoom control to examine the molecule in greater detail.
1. How many strands does the molecule have?
The molecule has two strands of DNA
twisted to form the double-helix.

2. Refer to the Component Panel for an illustration of each of the bases that make up
DNA. How many different types of nucleotide bases make up the DNA molecule
in the DNA Viewer?

There are four different nucleotide bases in the

DNA molecule: adenine (A), thymine (T),
guanine (G) and cytosine (C).

3. The strands of DNA are held together by base pairs. Examine the DNA molecule for
instances of A.

Table 1: Base Pairs Highlight all of the base pairs you observe.
A with A T with A G with A C with A
A with T T with T G with T C with T
A with G T with G G with G C with G
A with C T with C G with C C with C

C. Click Play at the bottom of the DNA Viewer to begin the animation.
 Watch the animation closely.

Click Reset; then, use the Step control to move through the animation a second time and
answer the following questions.

4. What happens to the strands of the original DNA molecule as replication begins
and proceeds?
The strands separate and become unwound from each other as replication proceeds.

5. Which of the original strands of the DNA molecule serves as a template for
replication?

Both of the original strands serve as templates for replication.

6. How many nucleotides are added to each new strand of DNA at a time?
One nucleotide base is added to the end of the new strand at a time

7. Examine several instances of A on one of the strands. What nucleotide is added to

the new strand at each A?
The base thymine, or T, is added to the new strand at every place
there is an A on the original strand.

8. Describe what happens to the DNA once synthesis of the new strands has been
completed.
The daughter molecules wind up forming two identical double-helices.

D. Reset and Step through the replication process again. The two strands of DNA are
antiparallel (in other words, the strands run in opposite directions).

9. Scientists often refer to the DNA structure during the first step of replication as
the replication bubble.
 Imagine a vertical line running through the middle of the replication bubble.
Focus on either the right or left side.

Are the two new strands synthesized in the same direction? NO If not,
describe how they differ.
The two strands are synthesized in different directions. In addition, on one
strand, the synthesis of new DNA proceeds in a continuous fashion from the
starting point, while on the other strand, synthesis of new DNA occurs in a step-
wise fashion in the direction opposite of that on the first strand.
10. Now compare both sides of the replication bubble. Does DNA replication occur
the same way on both sides? Describe any differences.

Yes, replication occurs similarly on both sides of the replication bubble. The main
difference is that replication on one side of the replication bubble is rotated 180º
relative to replication on the other side.

E. Observe the replicated, daughter DNA molecules and answer the following questions.

11. Is the newly synthesized DNA strand identical to, complementary to, or unrelated
to the template? Explain.
Each newly synthesized strand is complementary to the old template strand.

12. Describe the DNA molecules after replication in terms of the strands that make
them up. Use the terms "old strand" when you refer to the original strands and
"new strand" when you refer to the newly synthesized strands.
Replication results in one double-stranded molecule becoming two double-stranded
molecules. Each of the "daughter" molecules contains one old strand and one new
strand.

F. Click Reset and then click on the A button in the Component Panel to remove A from the
replication process. Play and Step through the process again, watching the animation
closely. Follow the same procedure for T, G and C. Answer the following question based
on your observations.
 13. Can replication substitute a nucleotide for the missing nucleotide? Can replication
skip a base? Explain your answer.
In the absence of a nucleotide, synthesis stops when it reaches the first position
where the missing nucleotide would normally be added.

Part II: The Replication Machinery

In Part II, you will investigate the roles each of the replication proteins plays in DNA replication,
from separation of the two strands of the original DNA molecule, through the actual replication
process to the final "tying up of the loose ends."

Click on The Replication Machinery above the DNA Viewer.

Refer to the Component Panel on the left of the window to identify each of the structures
or components of DNA replication by symbol.

Click Play to begin the animation. Observe the animation and fill in the following table.

Component Predict a function for this Effect of Removing this

Component Component

Helicase In the absence of a nucleotide, Without helicase, the DNA

synthesis stops when it reaches cannot be unwound.
the first position where the
missing nucleotide would
normally be added.

SSB (single-strand DNA binding SSB binds to single-stranded DNA Without SSB, the separated
protein) and prevents the strands from strands of DNA come back
coming back together. together and rewind before
replication can occur.

Primer The primer serves as the starting Without primers, new synthesis
point for the synthesis of new of DNA cannot begin.
DNA.

DNA Polymerase III Pol III synthesizes new strands of Without DNA polymerase III,
DNA using the old strands as DNA replication cannot occur.
templates.
DNA Polymerase I Pol I removes the primers and Without DNA polymerase I, the
fills in the gaps with newly primer cannot be removed and
synthesized DNA. replaced with DNA.

Ligase Ligase joins the ends of the Without ligase, the newly
newly synthesized DNA synthesized DNA fragments
fragments to form one cannot be joined into one
continuous strand. continuous strand.

Critical Thinking:

Many of the drugs doctors use to treat cancer or diseases, such as AIDS, are effective because the
drugs specifically inhibit DNA replication.

Why might these drugs cause hazardous side effects for people taking them?

These drugs would also be expected to inhibit replication of the DNA needed for division
of normal healthy cells.

In spite of the side effects, these drugs continue to be effectively used to combat disease.
What might this observation tell you about the diseases themselves?
The usefulness of replication inhibitors to treat diseases, such as cancer and AIDS, suggests
that these diseases are especially dependent on DNA replication for their own
progression.

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