The three-dimensional structure of DNA:

Replicating photograph 51
How do we know that the DNA molecule is a double helix?

Goal: In this experiment, students will reproduce the experiment done by Rosalind Franklin that
showed that the structure of the DNA molecule is a double helix. However, the students will be
using light rather than x-rays and a helical tungsten filament rather than DNA.
Background:

One of the most useful ways to understand how something works is to look at its
structure. This is why the discovery of the helical structure of DNA started a DNA
revolution that would last the next 50 years. Knowing the structure of DNA allowed us to
understand how genes work and how they are replicated from one cell to another for
generations. It allowed us to better understand inherited diseases and how one single
change or mutation could lead to those diseases. Knowing the structure of DNA lead to
DNA fingerprinting, which has revolutionized forensic sciences by matching DNA
samples to crime suspects, and can also be used to determine the paternity or ancestry
of humans or pets. Recently, new developments in DNA technology (including cutting
edge CRISPR technology) could lead to gene editing and gene therapies. But, this all
started with determining the structure of DNA. Back in the 1800’s, scientists knew about
genetic traits, but it wasn't until 1943 that they learned that DNA was the "genetic factor"
and not until 1953 that the helical structure of DNA that we know today was described.

While the discovery of the structure of DNA involved four scientists, many scientific
breakthroughs had to occur for the structure of DNA to be found. Some of these include:
1) the progress made by X-ray crystallographers in studying organic macromolecules;
2) the growing evidence supplied by geneticists that it was DNA, not protein, in
chromosomes that was responsible for heredity; 3) Erwin Chargaff’s experimental
finding that there are equal numbers of A and T bases and of G and C bases in DNA;
and 4) Linus Pauling’s discovery that the molecules of some proteins have helical
shapes—arrived at through the use of atomic models and a keen knowledge of the
possible disposition of various atoms. However, the most important scientific
breakthrough was a photograph, photograph 51, which was taken by Rosalind Franklin
and her student.

In 1951, Franklin was offered a 3-year research scholarship at King's College in London
to set up and improve the x-ray crystallography unit there. X-ray crystallography is a
biophysical technique where scientists make crystals of molecules and shine x-rays on
them. The x-rays diffract and the diffracted pattern can be used to reconstruct the
molecule’s structure. DNA is too small to be seen with the naked eye, but using x-ray
crystallography, we can determine the positions of all the atoms in the DNA, and
thereby generate a 3D model based on those observations.

When Franklin arrived at King’s College, Maurice Wilkins was already using X-ray
crystallography to try to solve the structure of DNA. It turned out that Franklin arrived
while Wilkins was away, and on his return, Wilkins assumed that she was hired to be his
assistant. It was a bad start to a relationship that never got any better. However,
pushing forward, Franklin and her student, Raymond Gosling, were able to use X-ray
crystallography to get high-resolution photographs of crystallized DNA fibers, including
photograph 51. From photograph 51, she deduced the basic dimensions of the DNA
strand and that it was a helix.
She presented her data at a lecture in King's College at which James Watson was in
attendance. Watson admitted to not paying attention at Franklin's talk and not being
able to fully describe the lecture and the results to Francis Crick. Watson and Crick had
been working on solving the DNA structure. Franklin did not know Watson and Crick as
well as Wilkins did and never truly collaborated with them. It was Wilkins who showed
Watson and Crick the X-ray data that Franklin had obtained. The data confirmed the 3-
D structure that Watson and Crick had theorized for DNA. In 1953, both Wilkins and
Franklin published papers on their X-ray data in the same Nature issue with Watson and
Crick's paper on the structure of DNA.

Figure 1 Top: a picture of Rosalind Franklin and Maurice Wilkins. Bottom left to
right: Picture of Watson and Crick with their 3D model of DNA, The first X-ray
diffraction picture of DNA and Rosalind Franklin's DNA Photograph 51.

In this experiment, you will use some household items to reproduce the photograph that
launched a DNA revolution, photograph 51. However, instead of a DNA molecule, you
will use a filament from an incandescent light bulb, which has a helical shape. Instead of
x-rays, you will shine visible light from a laser pointer on the filament. Looking at the
diffraction pattern of the filament on a wall, you will be able to see the characteristic
“cross pattern” that is also shown in photograph 51

Parts List
● A small helical structure (I got mine from an old incandescent light bulb)
● Laser pointer (make sure it is not and LED one, I got mine from a pet store)
● White paper
Notes:Tungsten filament from the light bulb is helical just like DNA.
The laser pointer will act as our X-ray beam.
The white paper will be our x-ray film.

Directions
Safety Precautions
● Even though it is a laser pointer and not an x-ray beam, avoid direct exposure to
the beam
● Do not stare into the beam
● Do not view the beam through optical instruments (this includes your camera
phone)

Experiment
1. Make sure that your laser is aligned with the filament.
2. Use a leveled surface for support.
3. Shine the laser into the tungsten filament as shown in the picture below:

4. Try to get a picture similar to the one below (you can dim the lights in the room):

5. Have a partner carefully draw the pattern on the paper. It does not have to be
perfect.
6. Scientists measure the space in between the spots to calculate the size of the helix.
This is why a good look at photograph 51 gave away the structure! They had already
since helical structures before (they knew helical proteins). The spacing in between
 the spots could be measured and it gave away that the there would be no steric
constraints if the phosphates were on the outside.

7. What it should look like

Extras - other things you could do

You can find other patterns to explore and learn. Try other shapes and see what you
can learn from the different diffraction patterns. You can see more diffraction pattern
calculations here: https://www.rigaku.com/downloads/journal/RJ32-
2/Rigaku%20Journal%2032-2_35-43.pdf