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A2.

15S

Student

Activity 2.15 Meselson and Stahls experiment


on DNA replication
Purpose
To describe the method of DNA replication by considering the different theories originally
proposed to explain the process, and the evidence which supports the theory that is now
accepted.

Procedure
Complete the interactive tutorial associated with the activity, and then complete this worksheet.
Matthew Meselson and Frank Stahl worked at the California Institute of Technology. In 1958 they
grew bacteria in growth medium containing ammonium ions (NH4+) as the source of nitrogen. The
type of DNA made by the cells depended on the type of nitrogen present in the bacterias growth
medium. They used two isotopes of nitrogen 14N and 15N. 14N is the common, light form (isotope)
of nitrogen. 15N is the heavier form. They then extracted DNA from the bacterial cells and centrifuged
the resulting solution to isolate the DNA. The DNA made with 14N and the DNA made with 15N
accumulated at different levels in the centrifuged solutions, according to the DNAs density.

Questions
Q1 In each of the label boxes in Figure 1 below, fill in the number to show the type of nitrogen present in
each band.
Q2 In each magnified circle in Figure 1, colour in the sections of DNA molecule found in each band to show
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the type of DNA in each band. Use your own colour code, selecting one colour for DNA made with N
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and another colour for DNA made with N. You will need a colour for medium DNA later, so red
(heavy), blue (light) and purple (medium DNA) would work.

Figure 1 Position of labelled nitrogen bands in the centrifuge tubes.

Q3 How did Meselson and Stahl produce bacterial cells containing only DNA
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made with N?
Bacteria containing DNA made with 15N were allowed to divide once in a solution
containing only 14N. Any new DNA made would contain 14N. After a single
replication the DNA was extracted and centrifuged. Figure 2 DNA found after
first replication.

Salters-Nuffield Advanced Biology, Pearson Education Ltd 2008. University of York Science Education Group.
This sheet may have been altered from the original. 1 of 3
A2.15S

Activity 2.15 Meselson and Stahls experiment on DNA replication Student

Q4 Colour in the band(s) in the centrifuge tube in Figure 2 to show


the density of the DNA that Meselson and Stahl found after this
first replication. Use your colour codes for heavy, medium and
light DNA.

Q5 Figure 3 shows DNA replication according to the theory of


conservative replication, Explain why the bands found by
Meselson and Stahl after one replication (shown in Figure 2)
refute the theory of conservative replication.
Bacteria containing DNA made with 15N were allowed to
divide twice in a solution containing only 14N. Any new DNA
made would contain 14N. After two replications the DNA was
extracted and centrifuged.
Q6 Figure 4 shows DNA replication according to the dispersive
14 15
theory, where new ( N) and original ( N) DNA are dispersed
throughout any new DNA molecules synthesised. This is theory 1
in the Interactive tutorial.
Figure 2 Conservative replication.

i In Figure 4, colour in the DNA and DNA nucleotides using your


colour code from question 2.
ii Now draw in the four DNA molecules produced when the DNA replicates for the second time, according
to the dispersive theory. Use your colour code for this.

Figure 4 The dispersive theory for replication.

iii Draw bands on the centrifuge tubes in Figure 5 to show the DNA present after the first and second
dispersive replications. Assume all the DNA has replicated after each cell division.

Figure 5 Bands that would occur after dispersive replication.

Salters-Nuffield Advanced Biology, Pearson Education Ltd 2008. University of York Science Education Group.
This sheet may have been altered from the original. 2 of 3
A2.15S

Activity 2.15 Meselson and Stahls experiment on DNA replication Student

b i In Figure 6 showing the semi-conservative theory of replication colour in the DNA and DNA
nucleotides using your colour code from question 2.
ii Now draw in the four DNA molecules produced when the DNA replicates for the second time,
according to the semi-conservative theory. This is where new DNA molecules have one strand of the
original (15N) DNA and one strand of the (14N) DNA. This is theory 2 in the interactive tutorial. Use
your colour code from question 2.

Figure 6 The semi-conservative theory for replication.

iii Draw bands on the centrifuge tubes in Figure 7 to show the DNA present after each replication.
Assume all the DNA has replicated after each cell division.

Figure 7 Bands that would occur after each replication.

Q7 Meselson and Stahl found equal amounts of light and intermediate density (medium) DNA present after
two DNA replications. Explain which of these three theories for DNA replication is supported by this
evidence and which is refuted. Use a separate sheet of paper if you need more space for your answer.

Salters-Nuffield Advanced Biology, Pearson Education Ltd 2008. University of York Science Education Group.
This sheet may have been altered from the original. 3 of 3