DNA: Definition, Structure, Discovery, Importance and

Denaturation

The structure of DNA and RNA. DNA is a double helix, while RNA is a single helix. Both have sets of nucleotides that
contain genetic information.

Credit: udaix Shutterstock

Definition
DNA is a complex, long-chained molecule that contains the genetic blueprint for building and
maintaining all living organisms. Found in nearly all cells, DNA carries the instructions needed to
create proteins, specific molecules essential to the development and functioning of the body. It
also transfers hereditary information between generations.

Deoxyribonucleic acid or DNA is a molecule that contains the instructions an organism needs to
develop, live and reproduce. These instructions are found inside every cell, and are passed down
from parents to their children.
DNA structure

Representations of the double-helical structure of DNA have become common in the popular
press as well as in the scientific literature. When the double helix was proposed by James Watson
and Francis Crick in 1953, it touched off a flood of research activity, leading to great advances in
molecular technology.

DNA is made up of molecules called nucleotides. Each nucleotide contains a phosphate group, a
sugar group and a nitrogen base. The four types of nitrogen bases are adenine (A), thymine (T),
guanine (G) and cytosine (C). The order of these bases is what determines DNA's instructions,
or genetic code. Human DNA has around 3 billion bases, and more than 99 percent of those
bases are the same in all people, according to the U.S. National Library of Medicine (NLM).

Similar to the way the order of letters in the alphabet can be used to form a word, the order of
nitrogen bases in a DNA sequence forms genes, which in the language of the cell, tells cells how
to make proteins. Another type of nucleic acid, ribonucleic acid, or RNA, translates genetic
information from DNA into proteins.

Nucleotides are attached together to form two long strands that spiral to create a structure called
a double helix. If you think of the double helix structure as a ladder, the phosphate and sugar
molecules would be the sides, while the bases would be the rungs. The bases on one strand pair
with the bases on another strand: adenine pairs with thymine, and guanine pairs with cytosine.

DNA molecules are long — so long, in fact, that they can't fit into cells without the right packaging.
To fit inside cells, DNA is coiled tightly to form structures we call chromosomes. Each
chromosome contains a single DNA molecule. Humans have 23 pairs of chromosomes, which
are found inside the cell's nucleus.

DNA discovery

DNA was first observed by a German biochemist named Frederich Miescher in 1869. But for
many years, researchers did not realize the importance of this molecule. It was not until 1953 that
James Watson, Francis Crick, Maurice Wilkins and Rosalind Franklin figured out the structure of
DNA — a double helix — which they realized could carry biological information.

Watson, Crick and Wilkins were awarded the Nobel Prize in Medicine in 1962 "for their
discoveries concerning the molecular structure of nucleic acids and its significance for information
transfer in living material." Franklin was not included in the award, although her work was integral
to the research. [Related: Unraveling the Human Genome: 6 Molecular Milestones]

Importance of DNA

DNA is central to biotechnology and medicine by virtue of the fact that it not only provides the
basic blueprint for all life, it is a fundamental determinant of how the body functions and the
disease process. Understanding the structure and function of DNA has helped revolutionise the
investigation of disease pathways, assess an individual’s genetic susceptibility to specific
diseases, diagnose genetic disorders, and formulate new drugs. It is also critical to the
identification of pathogens.

Aside from its medical uses, the fact that DNA is unique to each individual makes it a vital forensic
tool identifying criminals, the remains of a missing person, and determining the biological parent
of a child. Within agriculture DNA is also used to help improve animal livestock and plants.

DNA sequencing

DNA sequencing is technology that allows researchers to determine the order of bases in a DNA
sequence. The technology can be used to determine the order of bases in genes, chromosomes,
or an entire genome. In 2000, researchers completed the first full sequence of the human genome,
according to a report by the National Human Genome Research Institute.

DNA testing

A person's DNA contains information about their heritage, and can sometimes reveal whether
they are at risk for certain diseases. DNA tests, or genetic tests, are used for a variety of reasons,
including to diagnose genetic disorders, to determine whether a person is a carrier of a genetic
mutation that they could pass on to their children, and to examine whether a person is at risk for
a genetic disease. For instance, mutations in the BRCA1 and BRCA2 genes are known to
increase the risk of breast and ovarian cancer, and analysis of these genes in a genetic test can
reveal whether a person has these mutations.
Genetic test results can have implications for a person's health, and the tests are often provided
along with genetic counseling to help individuals understand the results and consequences of the
test.

There are now many at-home genetic testing kits, but some of them are unreliable. Also, NBC
News reports that people should be careful with these kits, since the tests are essentially handing
over a person's genetic code to a stranger.

Denaturation of DNA

We have already seen that the hydrogen bonds between base pairs are an important factor in
holding the double helix together. The amount of stabilizing energy associated with the hydrogen
bonds is not great, but the hydrogen bonds hold the two polynucleotide chains in the proper
alignment. However, the stacking of the bases in the native conformation of DNA contributes the
largest part of the stabilization energy. Energy must be added to a sample of DNA to break the
hydrogen bonds and to disrupt the stacking interactions. This is usually carried out by heating the
DNA in solution.

The heat denaturation of DNA, also called melting, can be monitored experimentally by observing
the absorption of ultraviolet light. The bases absorb light in the 260-nm-wavelength region. As the
DNA is heated and the strands separate, the wavelength of absorption does not change, but the
amount of light absorbed increases. This effect is called hyperchromicity. It is based on the fact
that the bases, which are stacked on top of one another in native DNA, become unstacked as the
DNA is denatured.

Because the bases interact differently in the stacked and unstacked orientations, their absorbance
changes. Heat denaturation is a way to obtain singlestranded DNA, which has many uses. When
DNA is replicated, it first becomes single-stranded so that the complementary bases can be
aligned. This same principle is seen during a chemical reaction used to determine the DNA
sequence. A most ambitious example of this reaction is described in the following Biochemical
Connections box.

Under a given set of conditions, there is a characteristic midpoint of the melting curve (the
transition temperature, or melting temperature, written Tm) for DNA from each distinct source.
The underlying reason for this property is that each type of DNA has a given, well-defined base
composition. A G–C base pair has three hydrogen bonds, and an A–T base pair has only two.
The higher the percentage of G–C base pairs, the higher the melting temperature of a DNA
molecule. In addition to the effect of the base pairs, G–C pairs are more hydrophobic than A–T
pairs, so they stack better, which also affects the melting curve.

Renaturation of denatured DNA is possible on slow cooling. The separated strands can
recombine and form the same base pairs responsible for maintaining the double helix.

New research on DNA

DNA research has lead to some interesting, and important findings in the last few years. For
example, a 2017 study published in the journal Science found that random mistakes in DNA, not
heredity or environmental factors, accounts for two-thirds of cancer mutations in cells.

Another 2017 breakthrough is the first sequencing of DNA from Egyptian mummies. "We were
excited to have at hand the first genome-wide data of ancient Egyptian mummies," said Stephan
Schiffels, leader of the Population Genetics Group at the Max Planck Institute for the Science of
Human History, in Jena, Germany. The findings were published in the May 2017 issue of the
journal Nature Communications.
Experiment: DNA Extraction from Fruit

DNA is the hereditary material in all cells. The experiment described below allows you to
isolate DNA from fruit.

To understand the important principles behind this step, it is not essential to do this experiment
at this stage or, in fact, at all. If you are not able to carry out this experiment, read through the
details below and try to work out why the procedure works in the way it does - a few questions
have been posted at the end of the method to help you think about this. You could set up a
discussion about the principles behind the experiment using the Comments boxes.

If you are studying this course with other people (e.g. within a school class), you might want to
think about doing the experiment together

If you do plan to carry out this experiment, you will need to read the instructions carefully and do
some planning before starting it.

Materials required
For this experiment you will need:

 Fruit – Kiwi, Strawberries, and Banana all work well

 5 g washing up liquid

 2 g salt

 100 ml tap water

 100 ml of ice cold alcohol (isopropyl alcohol can usually be found at the pharmacists); put
in a freezer for at least 30 mins before starting the experiment

 Access to hot water - about 60 °C

 Sieve or coffee filter paper

 Two glass beakers (or old jam-jars)

 Several bowls of different sizes, including a large bowl for making a water bath
  A paperclip

 Safety spectacles - if desired

Experiment to purify DNA from fruit

Step 1: Mash up the fruit of your choice in a bowl. Bananas, kiwis and strawberries all work well.
(Remove the skin of the bananas and kiwi, we just want the insides!)

Step 2: In a separate bowl, mix the washing up liquid, salt and tap water. Stir gently trying to avoid
making too many bubbles in the mixture. This is your extraction buffer.

Step 3: Add the fruit to the extraction buffer and mix again. Mash your fruit sample as much as
you can, but again, try to avoid making too many bubbles.

Step 4: Make a water bath with a temperature of about 60 °C. (A large washing up bowl works
well for this.) Leave the fruit extraction mixture to incubate for 15 minutes.

Step 5: After 15 minutes, filter your fruit mixture through a fine sieve or coffee filter. This will
remove all the solid material that you don’t want. You should be left with a clear(ish) liquid.

Step 6: Take the ice cold alcohol and very slowly, drop by drop, pour it down the inside of the
container with your fruit mixture. What you want to do is produce a layer of the alcohol floating on
top of the fruit mixture.

Step 7: At the interface between the alcohol and the fruit mixture, you should see a white cloud-
like substance forming. Use a hook (a bent paperclip would work) to slowly draw the DNA up and
out of the solution.

1. Look at each step of the protocol. What are the different steps and reagents (washing-up
liquid, salt, water, alcohol) used for in the experiment?

2. How are you able to see the DNA?

3. Do you think that the sample you have prepared is “pure DNA”?
Urine

Urine, liquid or semisolid solution of metabolic wastes and certain other, often toxic, substances
that the excretory organs withdraw from the circulatory fluids and expel from the body.
The composition of urine tends to mirror the water needs of the organism. Freshwater animals
usually excrete very dilute urine. Marine animals tend to combat water loss to their
salty environment by excreting concentrated urine; some develop methods actively to expel salts.
Terrestrial animals, depending on their habitat, usually retain water and secrete a highly
concentrated urine.

In most mammals, including humans, the formation of urine begins in the nephrons of the kidneys
by filtration of blood plasma into the nephron; the fluid found within the nephron is essentially the
same as blood plasma without the macromolecules (e.g., proteins). As the fluid passes along the
nephron tube, water and useful plasma components such as amino acids, glucose, and other
nutrients are reabsorbed into the bloodstream, leaving a concentrated solution of waste material
called final, or bladder, urine. It consists of water, urea (from amino acid metabolism), inorganic
salts, creatinine, ammonia, and pigmented products of blood breakdown, one of which
(urochrome) gives urine its typically yellowish colour. In addition, any unusual substances for
which there is no mechanism of reabsorption into the blood remain in the urine. The products
of nucleic acid breakdown are present as allantoin in most mammals and as uric acid in man and,
through a quirk of breeding, in the Dalmatian dog.

asparagus; urineLearn why eating asparagus can change the smell of urine.© American
Chemical Society

urine; alchemyLearn about the history of urine in alchemy and chemical experiments.© American
Chemical Society
In most birds, reptiles, and terrestrial insects, the end product of amino acid metabolism is not
water-soluble urea but insoluble uric acid. The urine of birds and reptiles is a whitish, aqueous
suspension of uric acid crystals that is passed into the cloaca and mixed with fecal material before
being expelled. The urine of terrestrial insects is solid and in some cases is stored as pigment in
the body rather than being expelled.

Amphibians and fishes excrete aqueous solutions of urea; unlike those of mammals, however,
their excretory organs do not reabsorb large quantities of water, so their urine remains dilute.
Some marine animals retain much of the urea in the blood, thus retarding osmotic water loss.

In small, primitive animals (teleost fishes, echinoderms, coelenterates, and single-celled animals),
particularly those that live in aqueous environments, the end product of amino acid metabolism is
the highly toxic gas ammonia, which is collected and expelled in a dilute aqueous solution. Many
of the smaller animals develop no excretory system; each individual cell disposes of its waste
products to the circulatory fluids, and the wastes then diffuse to the surrounding medium.

Urine is a liquid byproduct of the body secreted by the kidneys through a process called urination
and excreted through the urethra. The normal chemical composition of urine is mainly water
content, but it also includes nitrogenous molecules, such as urea, as well as creatinine and other
metabolic waste components. Other substances may be excreted in urine due to injury
or infection of the glomeruli of the kidneys, which can alter the ability of the nephron to reabsorb
or filter the different components of blood plasma.

Normal Chemical Composition of Urine

Urine is an aqueous solution of greater than 95% water, with a minimum of these remaining
constituents, in order of decreasing concentration:

 Urea 9.3 g/L.

 Chloride 1.87 g/L.
 Sodium 1.17 g/L.
 Potassium 0.750 g/L.
 Creatinine 0.670 g/L .
  Other dissolved ions, inorganic and organic compounds (proteins, hormones,
metabolites).

Urine is sterile until it reaches the urethra, where epithelial cells lining the urethra are colonized
by facultatively anaerobic gram-negative rods and cocci. Urea is essentially a processed form of
ammonia that is non-toxic to mammals, unlike ammonia, which can be highly toxic. It is processed
from ammonia and carbon dioxide in the liver.

ABNORMAL TYPES OF URINE

There are several conditions that can cause abnormal components to be excreted in urine or
present as abnormal characteristics of urine. They are mostly referred to by the suffix -uria. Some
of the more common types of abnormal urine include:

 Proteinuria—Protein content in urine, often due to leaky or damaged glomeruli.

 Oliguria—An abnormally small amount of urine, often due to shock or kidney damage.
 Polyuria—An abnormally large amount of urine, often caused by diabetes.
 Dysuria—Painful or uncomfortable urination, often from urinary tract infections.
 Hematuria—Red blood cells in urine, from infection or injury.

 Glycosuria—Glucose in urine, due to excess plasma glucose in diabetes, beyond the

amount able to be reabsorbed in the proximal convoluted tubule.

Urine Formation in the Human Body

The kidneys filter unwanted substances from the blood and produce urine to excrete them. There
are three main steps of urine formation: glomerular filtration, reabsorption, and secretion. These
processes ensure that only waste and excess water are removed from the body.

The Glomerulus Filters Water and Other Substances from the Bloodstream

Each kidney contains over 1 million tiny structures

called nephrons. Each nephron has a glomerulus, the
site of blood filtration. The glomerulus is a network of
capillaries surrounded by a cuplike structure, the
glomerular capsule (or Bowman’s capsule). As blood
flows through the glomerulus, blood pressure pushes
water and solutes from the capillaries into the capsule
through a filtration membrane. This glomerular filtration begins the urine formation process.

The Filtration Membrane Keeps Blood Cells and Large Proteins in the Bloodstream

Inside the glomerulus, blood pressure pushes fluid

from capillaries into the glomerular capsule through a
specialized layer of cells. This layer, the filtration
membrane, allows water and small solutes to pass
but blocks blood cells and large proteins. Those
components remain in the bloodstream. The filtrate
(the fluid that has passed through the membrane)
flows from the glomerular capsule further into the
nephron.

Reabsorption Moves Nutrients and Water Back

into the Bloodstream

The glomerulus filters water and small solutes out of the

bloodstream. The resulting filtrate contains waste, but also
other substances the body needs: essential ions, glucose,
amino acids, and smaller proteins. When the filtrate exits
the glomerulus, it flows into a duct in the nephron called
the renal tubule. As it moves, the needed substances and
some water are reabsorbed through the tube wall into adjacent capillaries. This reabsorption of
vital nutrients from the filtrate is the second step in urine creation.

Waste Ions and Hydrogen Ions Secreted from the Blood Complete the Formation
of Urine

The filtrate absorbed in the glomerulus flows through

the renal tubule, where nutrients and water are
reabsorbed into capillaries. At the same time, waste
ions and hydrogen ions pass from the capillaries into
the renal tubule. This process is called secretion.
The secreted ions combine with the remaining filtrate
and become urine. The urine flows out of the nephron tubule into a collecting duct. It passes out
of the kidney through the renal pelvis, into the ureter, and down to the bladder.

Urine Is 95% Water

The nephrons of the kidneys process blood and create urine

through a process of filtration, reabsorption, and secretion.
Urine is about 95% water and 5% waste products.
Nitrogenous wastes excreted in urine include urea,
creatinine, ammonia, and uric acid. Ions such as sodium,
potassium, hydrogen, and calcium are also excreted.

Urinalysis: An Example of Urine Test

Urinalysis is a series of tests on your pee. Doctors use it to check for signs of common conditions or
diseases. Other names for it are urine test, urine analysis, and UA.
You may have a urinalysis as part of a routine check of your overall health, for instance as part of an
annual physical. Urinalysis is one way to find certain illnesses in their earlier stages. They include:
 Kidney disease
 Liver disease
 Diabetes
Your doctor may also want to test your pee if you’re getting ready to have surgery or are about to be
admitted to the hospital. Urinalysis can be part of a pregnancy checkup, too.
If you have symptoms of a kidney or urinary tract problem, you may have the tests to help find out
what the problem is. Those symptoms include:
 Pain in your belly
 Pain in your back
 Pain when you pee or needing to go frequently
 Blood in your pee
How Does It Work?
There are three ways to analyze urine, and your test might use all of them.
One is a visual exam, which checks the color and clarity. If your pee has blood in it, it might be red or
dark brown. Foam can be a sign of kidney disease, while cloudy urine may mean you have an
infection.
A microscopic exam checks for things too small to be seen otherwise. Some of the things that
shouldn’t be in your urine that a microscope can find include:
 Red blood cells
 White blood cells
 Bacteria
 Crystals (clumps of minerals – a possible sign of kidney stones)
The third part of urinalysis is the dipstick test, which uses a thin plastic strip treated with chemicals.
It’s dipped into your urine, and the chemicals on the stick react and change color if levels are above
normal. Things the dipstick test can check for include:
 Acidity, or pH. If the acid is abnormal, you could have kidney stones, a urinary tract
infection (UTI) or another condition.
 Protein. This can be a sign your kidneys are not working right. Kidneys filter waste products
out of your blood.
 Glucose. A high sugar content is a marker for diabetes.
 White blood cells. These are a sign of infection or inflammation, either in the kidneys or
anywhere else along urinary tract.
 Nitrites. This means that there is an infection with certain kinds of bacteria.
 Bilirubin. If this waste product, which is normally eliminated by your liver, shows up, it may
mean your liver isn’t working properly.
 Blood in your urine. Sometimes this is a sign of infections or certain illnesses.