DNA Fingerprinting Definition

Every human being has a unique set of fingerprints which can be used for identification.
Historically, this was especially helpful when attempting to solve a crime. Fingerprints
detected at the crime scene would be compared and hopefully matched to criminals in a
fingerprint database. After the discovery of DNA and along with advances in technology,
another method of identification besides fingerprinting was developed called DNA
fingerprinting. DNA Fingerprinting can be defined as a method of using DNA to create a
unique genetic pattern for every person. For example, if any cells are left behind at a crime
scene, DNA can be obtained from those cells and put through a series of steps that will
ultimately create a unique pattern for that individual. This unique pattern is what is referred to
as the DNA fingerprint. Like traditional fingerprinting, this DNA fingerprint is then
compared to a known sample to see if there is a match.

This image demonstrates what a DNA Fingerprint looks like. Observe the bands which
create unique patterns.

Another term that is used synonymously with DNA fingerprinting is DNA profiling. Like
DNA fingerprinting, DNA profiling is also a technique that identifies patterns of DNA
sequences that are unique to an individual. The unique patterns created from DNA profiling
can also be used to identify human remains, determine paternity, or compare two species for
genetic similarities.
DNA can be taken from any cell in the body, including liquid tissue such as blood. DNA is
often left behind in small amounts at a crime scene, unbeknownst to the criminal. DNA can
also be collected from hair follicles if hair has been forcibly removed, from skin cells or
saliva left on a cigarette butt, or from fingernail scrapings of a victim.
Although no two people have the same DNA fingerprint (except for identical twins), DNA
fingerprinting often works best when eliminating a suspect to prove innocence. If the DNA
sample from a suspect does not match the crime scene sample, then it can be reasonably
concluded that person was not at the crime scene and thus did not commit the crime. A
matching sample, in contrast, simply means that the suspect was present at the crime scene at
some point in time and does not prove that the suspect committed the crime.
A great early example of the use of DNA fingerprinting to prove innocence is the case of
Kirk Bloodsworth, charged with child rape and murder in 1985 and put on death row.
Bloodsworth adamantly maintained that he was innocent, claiming he was charged based on
faulty eyewitness testimony. In 1993, Bloodsworth was exonerated by DNA fingerprinting
because the drops of semen that were left behind at the scene did not match his DNA. Many
other wrongfully convicted persons have been freed based on DNA evidence, which is
largely attributed to the Innocence Project that was founded in 1992.

Kirk Bloodsworth was exonerated of the rape and murder of a child because of DNA Fingerprinting.

How are DNA Profiles Stored?

DNA profiles, also called DNA types, are created through the process of gel electrophoresis.
Once created, these profiles are photographed and then stored in national databases. In 1994,
congress passed the DNA Identification Act which gave the FBI permission to begin storing
profiles of criminals (starting with sex offenders) in a database. This database is called
the National DNA Index System (NDIS) and has expanded to include all types of criminals
and suspects. This is particularly helpful to law enforcement agencies if a suspect crosses
state lines.

DNA Fingerprinting Steps

A very small amount of DNA is needed to create a DNA fingerprint. The following steps
explain this process:
Step 1 (DNA Extraction): DNA is isolated from the cells containing it through a process
called DNA extraction. During this process, the cell membrane and nuclear membrane will be
chemically broken down to allow the DNA to flow out. Additional chemical reactions are
performed to isolate and purify the DNA to get it ready for the next step.
Step 2 (PCR): Once the DNA is extracted, it is put through a process known as
a Polymerase Chain Reaction (PCR). In this process, the DNA sample is copied many
times over until there is an amount that is sufficient to create the DNA fingerprint.
Step 3 (Restriction Enzyme Treatment): The copies of DNA will be cut up or digested
using special molecular scissors known as restriction enzymes. These enzymes will cut
DNA at specific genetic sequences. Because each person has unique DNA, the places that the
restriction enzymes cut will be unique to each person. This will create fragments of DNA that
are different sizes. Each person will have fragments of different lengths. These fragments are
given the name of Short Tandem Repeats (STRs), referred to as such because most of our
DNA is made up of non-coding areas that have repeating sequences. These regions are
typically where the restriction enzymes will make their cuts.
Step 4 (Gel Electrophoresis): The digested DNA samples are loaded into wells that are
formed in a special gelatin called an agarose gel. This porous gel has the consistency of Jell-
O, allowing the DNA fragments to move through openings in it. An electric current is applied
to the gel to move the DNA fragments through. Because DNA has a negative charge to it, the
side of the gel where the DNA is loaded will have a negative charge applied to it. The
opposite side of the gel will have a positive charge applied. The negatively charged DNA will
be pulled toward the positive side of the gel through the pores. The larger fragments of DNA
will not travel as far and will stay closer to the wells while the smaller fragments will travel
further. The DNA fragments will separate out over a period of time based on their size,
creating the unique pattern referred to as the DNA fingerprint.

DNA samples are loaded into wells in an agarose gel. An electric current is applied which
moves the fragments from the negative end toward the positive end.

Uses of DNA Fingerprinting

DNA fingerprinting started as a way to match a potential suspect(s) to a crime scene. In the
realm of forensics, if DNA at a crime scene is found to match a suspect, it can support other
evidence collected to establish a high probability of guilt. DNA fingerprinting is used not
only to assist in proving guilt; it can also exonerate the innocent if a suspect's DNA does not
match any samples taken from the scene. DNA fingerprinting has evolved into DNA
profiling, meaning there are now several uses of this technology:
  Paternity testing- A DNA profile of a child will have a 50% match to the mother and
50% to the father. If a father is unknown, a DNA profile can be created for potential
fathers and if the profile has a 50% match to the child, paternity is established.
 Victim identification- Plane crashes and fires are two examples of traumatic
incidents that often leave bodies in unidentifiable conditions. Thankfully, DNA can be
extracted to create a DNA profile that can then be compared to samples prepared from
victims. Samples could also be compared to a close relative of the victim to help
determine identity.
 Inheritance cases- Occasionally, a wealthy person dies and an unknown person
claiming to be a relative comes forward to collect an inheritance. In this case, DNA
profiling is useful to determine the relationship between the deceased and those
claiming to be closely related.
 Ancestry- In recent years, companies such as Ancestry.com have become popular to
assist individuals in finding out more about their genetic roots.
 Species comparisons- DNA profiles can also be created for non-human biological
species and is being used to conduct research regarding relationships of these species
or breeds within a species to each other. A familiar example might be using DNA
profiling to determine what percentage of dog breeds can be found in a mixed breed
where the parents are unknown.

Lesson Summary
DNA fingerprinting is a technique that involves isolating and extracting DNA from cells,
amplifying it through PCR, and running it through gel electrophoresis. Restriction
enzymes are used to cut the DNA into fragments of different sizes that are called short
tandem repeats or STRs. These different sized fragments will move through the gel when
electricity is applied, and they will separate out by size. This causes a pattern which will be
unique for every person and this is what is referred to as the DNA fingerprint.
DNA fingerprinting is also known as DNA profiling. A DNA profile can be used to
determine if a suspect was at a crime scene but is more often used to exonerate suspects if the
DNA fingerprint does not match crime scene samples. DNA profiles, also called DNA types,
are saved in national databases like the National DNA Index System (NDIS). Additional
uses of DNA profiling include paternity testing, inheritance cases, determining ancestry,
identifying unknown human remains, and even in species comparisons.

What Is DNA Fingerprinting?

Your DNA is very special because it is what makes you, you. Unless you have an identical
twin, no one else has the same DNA as you. This genetic identification comes from your
parents - half from mom and half from dad. But no matter how many kids your parents have,
your DNA will always be unique from everyone else's.
And this is what makes DNA fingerprinting such a useful tool: it allows us to identify an
individual from biological samples. It's called fingerprinting because, just like identifying an
individual based on the unique patterns we find on their fingers, we identify an individual
based on the unique fingerprint of their DNA. Because of this, you may have also heard it
called DNA profiling or identity testing.
For example, if you leave a hair behind somewhere, the DNA in that strand of hair can be
matched to you by taking a sample of DNA from another part of your body. As you can
imagine, this is really helpful for catching criminals if they leave DNA behind at a crime
scene. But we'll get into that a little later on.

How It Works
First, let's talk about how we go about creating the fingerprint. Again, pretty much all of your
cells contain your DNA so we can get a sample from bodily fluids like blood, but it can also
come from hair, bone, skin, and other types of cells.
The DNA is extracted from the sample. This process will use chemicals that break open the
cell to extract and isolate DNA from the nucleus. The DNA will then go through quantitation,
where the quality and quantity of the DNA is assessed. The DNA is then augmented using a
technique called _PCR_, or _polymerase chain reaction_, which replicates and amplifies the
specific site of DNA that will be analyzed. This is necessary because there's usually only a
small amount of DNA available in the reference sample, and this step lets us copy the DNA
to get enough to make a profile for the individual.
This amplified DNA is then cut at specific sequences with restriction endonucleases, and this
is where the magic happens. Because each person's DNA is different, these enzymes will cut
it at different sites, leaving us with pieces of different sizes. Specifically, these regions are
called STRs, or short tandem repeats, which are non-coding DNA regions with specific and
repeated nucleotide sequences. The number of times the sequence is repeated is unique to
each individual, giving each of us fragments of different lengths. During this process, primers
are used that will attach a fluorescent tag to the fragments.
But we're not quite done yet! The next step is to separate the fragments based on their size
through gel electrophoresis. That sounds like a mouthful, but it's actually pretty simple. The
DNA fragments are put into agarose gel (which is a lot like gelatin), and then an electric
current is applied to the gel. The shorter fragments move to the positive pole more quickly
than the longer fragments, and the pattern of the fragment separation is compared to the
reference sample for a match. We can see the different fragments because they've been
stained with a fluorescent dye, and this visualization in the gel is called electrophoresis
visualization.
The next step is Southern Blotting, where the separated DNA is transferred to a solid matrix
or membrane, which allows for the fragments to be secured and detected by a radioactive
probe. An x-ray film is placed next to the solid matrix, which will detect the patterns from the
probe, making the bands visible. The bands, which look almost like a barcode, are the DNA
fingerprint or profile of the individual that the DNA belongs to. This DNA profile can be
compared to a suspected DNA profile or run through the CODIS database to search for a
match.

Uses of DNA Fingerprinting

This matching makes DNA fingerprinting very useful for criminal and forensic analysis. If a
criminal leaves behind a sample of DNA at the crime scene, then that sample can be
compared to the DNA of various suspects to determine who was there when the crime was
committed.
You see this on TV dramas all the time now, but the first time this technique was used to
solve a crime was only as recently as 1987. And DNA fingerprinting is not only used to
determine crimes that are committed now, but has also been used to identify criminals of past
crimes and prove the innocence of many people who were put in jail for crimes they never
actually committed.
Because you get your genes from your parents, your DNA is very similar to theirs as well.
This makes DNA fingerprinting useful for matching children with their parents. This could be
through paternity testing, which tests whether a man is the father of a certain child, but it
could also be to identify children who are lost during natural disasters such as earthquakes,
hurricanes, and tsunamis and reunite them with the correct family.
DNA fingerprinting can also be used to identify individuals who have died but are
unidentifiable - for example, identifying the victim of a crime or a natural disaster. In fact,
DNA fingerprinting was used during 9/11 to identify many of the victims whose bodies were
too damaged to identify otherwise.

Lesson Summary
Just like your fingerprints, your DNA is unique to you. It's because of this unique identifier
that DNA fingerprinting can be used to identify individuals from biological samples. This
may be a sample left behind at a crime scene, to determine paternity, or even to identify
victims of crimes or natural disasters.
To identify who the sample belongs to, DNA is extracted from the sample (which could be
blood, hair, skin, bone, or other tissue) and amplified through PCR, or polymerase chain
reaction. Once the DNA is replicated, it's cut at STRs, or short tandem repeats, by
restriction endonucleases that separate the DNA into fragments unique to the individual.
While we all have these nucleotide sequences found in STRs, the number of times the
sequence repeats is unique to each individual and, therefore, gives each of us fragments of
different lengths.
These fragments (which have been dyed during PCR) are then put into agarose gel that has an
electric current applied to it, a process called gel electrophoresis. The electric current
separates the unique fragments by size, and the pattern of how they separate can be compared
to an individual to determine a match. The final steps are the use of Southern Blotting and an
x-ray image to obtain the DNA profile.