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VLab: DNA Replication: Data Sheet

Table 1: Base Pairs

Circle 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

Figure 1: Components of DNA Replication

DNA Polymerase III

Single-strand Binding Proteins

DNA Polymerase I

Helicase

Primer

Ligase

Copyright © 2011, SAS Institute Inc., Cary, NC, USA, All Rights Reserved 3/1/2011
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Table 2: DNA Replication Components and Their

Functions

Component Predict a function for this Effect of Removing this Component

Component

Helicase Unwinds the DNA strand The DNA will not be unwound

SSB (single-strand DNA Prevents the DNA from rewinding by Allows the molecule to rewind before the
binding protein) stabilizing its strands replication can occur.

Primer Initiates DNA Synthesis New synthesis of DNA cannot begin.

DNA Polymerase III Replicates DNA DNA replication cannot occur.

The primer cannot be removed and

DNA Polymerase I Replaces RNA primer with DNA
replaced with DNA.

The newly synthesized DNA fragments

Allows the newly synthesized DNA
Ligase cannot be joined into one continuous
fragments to be joined together
strand.

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Copyright © 2009, SAS Institute Inc., Cary, NC, USA, All Rights Reserved 2/18/2009

Part I: Replicating DNA

1. Summarize the process of DNA replication. Be sure to describe the DNA before replication, each step
in the replication process and the DNA molecules after replication has finished.

 The DNA consists of 2 strands with 4 bases (Adenine, Guanine, Cytosine and Thymine. The
separation of the two single strands of DNA creates a ‘Y’ shape called a replication ‘fork’. The two
separated strands will act as templates for making the new strands of DNA. Next, each strand will
be synthesized in opposite directions. Once the synthesis is complete, the DNA ligase seals up the
sequence of DNA into two continuous double strands. The result of the replication is two DNA
molecules, each consisting of one new chain of nucleotides and one old chain of nucleotides.The
bases on each DNA chain are said to be complementary to each other.

2. Define the term complementarity using specific examples of which bases occur together.

 The base pair is partnered to only one other kind of base pair. So, A will always be partnered with
T (and vice versa) except for when its RNA and G will always be paired with C and vice-versa. So, if
one base pair is present, it’s automatic that the base pair to be partnered with it will automatically
be its complementary one.

3. Describe how the concept of complementarity affects our understanding of DNA replication.

 Because of the complimentary base pairs, each strand of the double helix can be used as a
template for the synthesis of the other strand.

4. A cell has run out of adenine (A) during replication of the following DNA. The open circle indicates the
position of the polymerase. Write out the sequence of bases that the cell can still complete on the
new strand.

 A mistake in DNA replication, in which the wrong base is placed in the growing DNA strand, can
have profound effects on the descendants of any cells that carry the mistake.

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5. In the following strand, the A has been changed to a G. Draw the DNA molecules that result from
replication of the altered strands.

G
T

C
A

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

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

 Because these drugs can also stop other normal healthy cells that need DNA replication, and
these cells need DNA replication in order for them to multiply and help the body work normally.

7. 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 diseases could be giving off something that makes the drugs target them more than the
body’s own cells.

Part II: The Replication Machinery

Compare your prediction for the role(s) that each component plays in DNA replication (Table 2, column 2)
with the effect you observed when each component was removed (Table 2, column 3) and answer the
following questions.

8. Did your predictions match your conclusions? Describe and explain any that did not.

 Yes, my predictions match my conclusions because their functions are basically the opposite of
the effects listed on column 3 when they are removed.

9. What component is needed to separate the strands in the original DNA molecule during the first step
in replication?

 Helicase

10. What keeps the separated DNA strands from coming back together?

 Single-strand binding proteins

11. What role do primers have in DNA replication?

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 They begin the process of DNA synthesis

12. What functions do DNA polymerases carry out?

 DNA Polymerase III begins forming the complementary strand of the DNA, while DNA Polymerase
I replace the RNA primers with DNA.

13. What step is required for completing the synthesis of a strand of DNA?

 The ends of the new DNA strands are joined together by ligase Recall your answers to questions 9
and 18 from Data & Observations.

14. Explain your observations, based on the fact that the two strands of DNA are antiparallel.

 DNA strands are antiparallel because the 3’ and 5’ ends run opposite from one another. This is to
allow the hydrogen bonds between the base pairs to exist (no unwinding would occur otherwise)
and to replicate the leading strand one way and the lagging strand in the opposite direction.

You are a crime scene investigator called in to help gather evidence at a murder scene. The only clues to
the identity of the murderer are some skin cells found beneath the victim's fingernails and a few hairs.
You know that human beings have unique DNA sequences that, like a fingerprint, can be used for
identification. Unfortunately, a relatively large amount of DNA is needed for DNA analysis, more DNA than
can be isolated from the samples you have collected. To get around this problem, you decide to use a
specialized application of DNA replication, called the polymerase chain reaction (PCR). In PCR, DNA is
isolated from skin cells, a blood smear, or even a few hairs and subjected to repeated rounds of
replication. Not all the DNA is copied, only those unique target sequences that can be used for
identification are replicated. With each round, the number of copies of target DNA doubles.

For these questions, assume that there was only 1 copy of target in the initial sample of DNA.

15. How many copies would there be after one round of PCR?

 2

16. How many copies would there be after 5 rounds of PCR?

 32

17. How many copies would there be after 10 rounds of PCR?

 1024

18. If 10 million copies of a target sequence were needed for analysis, how many rounds of PCR would be
necessary?

 24

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