Obtaining DNA evidence is only a small part of the equation for proving a person's guilt or innocence.

In
fact, any forensics analysis - from criminal evidence to resolving paternity disputes - has DNA as a mere
starting point. The true value lies in the outcome of DNA analysis, which can be performed through one or
more of many different techniques.

Polymerase Chain Reaction (PCR) Analysis

PCR analysis is a technique that allows technicians to create millions of precise DNA replications from a
single sample of DNA. In fact, DNA amplification alongside PCR can let forensic scientists perform DNA
analysis on samples that are as tiny as only a couple of skin cells. In contrast to some other DNA analysis
techniques, PCR analysis has the advantage of analysing minuscule sample sizes, even if they are
degraded although they must not be contaminated with DNA from other sources during the collection,
storage and transport of the sample.

Restriction Fragment Length Polymorphism (RFLP)

RFLP is a technique that is not widely used now but it was one of the first techniques used for DNA
analysis in forensic science. Large sample sizes are needed for RFLP relative to newer techniques -
usually a sample would need to be approximately the size of a one-pound coin. While that in itself may
sound small, it is large relative to other techniques such as PCR analysis that require only a few cells for
successful sequencing. In RFLP, the different lengths of DNA fragments are analysed. These fragments
are from the digestion of a sample of DNA with a restriction endonuclease enzyme. The enzyme chops
DNA in a certain style - the restriction endonuclease recognition site. Whether or not particular recognition
sites are present will provide different lengths of DNA fragments, which are then divided up through
electrophoresis. DNA probes then serve to hybridise the fragments through complementary binding.

Short Tandem Repeat (STR) Analysis

STR analysis works to examine individual areas in DNA. The differences from the collective areas of one
person to another can allow for distinguishing between individuals. In criminal investigations, there are
thirteen regions that are analysed and compared to establish profiles. In fact, DNA databases used at the
government level involve the sequence of these thirteen regions. The chances of two people having the
exact same thirteen regions is virtually impossible - likely one in a billion. A common DNA joke is that a
person's odds of winning the lottery are higher than finding a perfect match for the thirteen regions.

Mitochondrial DNA Analysis

Mitochondrial DNA analysis works well on samples that are unable to be analysed through RFLP or STR
analysis. There are two kinds of DNA in the cell - mitochondrial DNA and nuclear DNA. With other types
of analysis, nuclear DNA is removed from the sample but with mitochondrial DNA analysis, DNA is
removed from the cell's mitochondria. Sometimes, a sample can be old and will no longer have nuclear
material in the cell, which poses a problem for the other types of DNA analysis. With mitochondrial DNA
analysis, however, mitochondrial DNA can be removed, thus having important ramifications for cases that
were not solved over many years. This means that mitochondrial DNA analysis can be very valuable in
investigations for a missing person. Mitochondrial DNA will be the same from a woman to her daughter
because it is passed on from the egg cell.

Y-Chromosome Analysis
Since the Y chromosome passes from a male to his son, analysing genetic markers on a Y chromosome
can be of aid in identifying familial ties in males or for analysing any evidence entailing many males.
Another benefit of Y-chromosome analysis is to establish a family line over many generations.

There are other types of analysis but these are some of the main traditional and current methods used to
analyse DNA. No doubt, new techniques will be developed that will be even more rapid, successful and
cost-effective.

The acid phosphate test is among the most common and is even available via home testing
kits, which can be purchased online. The acid phosphate test detects a protein in semen
known as acid phosphatise. The test is referred to as a presumptive test because these tests
make use of a certain chemical to establish the presence of a certain body fluid or chemical.
In forensic science, presumptive tests can do two things: they either exclude a substance
from being semen or confirm with a good probability that the substance is semen. The acid
phosphate test hydrolyzes the protein in the semen sample; consequently, if semen is
actually present, a color change will be noted in the intensely colored azo dyestuff.
Presumptive tests have the chance of false positive results, which can indeed be misleading.
Further confirmatory testing is required following the results of a presumptive test that
gives a positive result.

The Rapid Stain Identification (RSID) is a confirmatory test. It is done by means of a kit
that contains an extraction/running buffer, water for swab wetting, swabs, scissors, transfer
pipettes, tube rack, protocols, and RSID-Semen strip tests. This test uses antibodies that
are specific for specific for human semenogelin on an immunochromatographic strip to
confirm the presence of a component of seminal fluid, Semenogelin. It should be carried out
whenever a presumptive test gives a positive result. The RSID test is very accurate and
specific for human semen testing and will even prove effective in diluted samples of semen,
semen mixed with other body fluids or the presence of fungi or bacteria in other words,
the semen test will not cross react. This is indeed an advantage as in cases of assault, cross
reactivity between similar proteins (body fluids containing similar components) occurs
frequently, and avoiding contamination between body fluids that may lead to inaccurate/
false positive or false negative results could pose problems.

Here is how it works:

1. First, it must be shown that the stain contains semen. Semen glows under ultraviolet light
and also changes color when exposed to specific chemicals.

2. If these tests are positive, the lab technicians cut a swatch of the fabric from the stain and
dissolve the organic matter in the stain by putting the swatch in a special solution.

3. The technicians then look for sperm cells in the solution with an ordinary low-power light
microscope. The sperm look like little lifeless tadpoles.

4. If there are visible sperm under the microscope, the techs next extract DNA from the sperm.
This is done with a mild detergent that bursts non-sperm cells. A rinse with water removes
broken cells. And then a stronger detergent is used to burst the sperm and recover their DNA.

5. The lab then compares the DNA from the semen with the suspect's DNA. A blood or saliva
sample from the suspect will supply enough DNA for the comparison.

6. If the stain is small and the amount of DNA minute, a method based on the polymerase chain
reaction, or PCR, is employed to make millions of copies of selected segments of the DNA.
Note that PCR does not change the DNA but merely amplifies the amount.

7. The DNA from the semen is then compared with the suspect's. This can be done using what
are called restriction fragment length polymorphisms, or RFLPs. To examine RFLPs, the DNA
from the sample is cut (restricted) by special enzymes. These restriction enzymes cut different
DNAs differently. If the restriction enzymes cuts the suspect's sample and the semen sample
into the same number of fragments of the same length, then the semen may be the suspect's.

8. The odds that the DNA in the semen is the suspect's are then calculated. The odds may, for
example, be one in 11 million that the DNA in the semen came from another man, not the
suspect.

The DNA tests of semen can take several days or several weeks before results are available,
depending upon which techniques have been used by the lab (and which lab has done the DNA
testing). DNA tests can show that the semen is not from the suspect. Or these tests can show that
the DNA in the sperm is not detectably different from the suspect's. But DNA testing can never prove
with complete certainty that the semen is from the suspect.

Proving a sexual assault with DNA

While the forensic testing of genetic material can resolve many legal situations, it is not
always a simple task. The tests performed by laboratories on DNA are only available to
identify a person or persons who have provided their samples for analysis. There is currently
no central database that contains all human beings genetic information. The reality is that
technology can provide a match only: meaning if there is a forensic sample and there is a
separate test performed on a suspect, the analysis can only tell if the two are a match or
not.

Recently, new options have surfaced to assist in identifying an unknown assailant. However,
its a small sample when compared to the population at large. In many places around the
world in the United States for instance a person charged or convicted with a crime
(depending on the state in question) will be forced to submit their DNA to a criminal
database. When a law enforcement agency is investigating a given sexual assault it is
possible that they can compare their forensic samples to the database in hopes of finding a
match. In other cases once there is a suspect found or identified, testing of this person can
be required in order to prove or disprove their involvement in the crime. This technology is
relatively new and laws protecting citizens from agencies looking at database DNA are
constantly evolving.

For how long can DNA forensic evidence be kept?

DNA is relatively stable. It is most likely that forensic samples collected from a rape victim
will yield results: however, time factors, chemical factors (such as washing using soaps and
detergents), external factors (such as temperature and humidity) and internal factors (other
bodily fluids) may affect the validity of a sample. The earlier samples are collected and
tested the higher the chances of yielding solid, reliable results. The following are just some
guidelines as to how long different DNA samples may remain viable:

DNA from fingers in vagina up to 12 hours.

DNA from a penis most likely to obtain a profile from the victim within the first 12
hours.
DNA from skin to skin contact (e.g., on bruises, or from kissing) can be detected up
to two days. This includes detection of body fluids, cellular material and lubricant. If
by chance, the person has not bathed or showered then the Forensic Science service
says that the relevant area can be swabbed up to 7 days after the incident.
Fingernail scrapings two days.
Oral (saliva and mouth swabs) two days.
Lubricant from a condom up to 30 hours.
Anal up to three days.
Vaginal up to seven days.
Fibers of anything put on the head can last up to seven days.
Semen can be detected on clothing despite washing.
In some cases of assault, the assailant may not have ejaculated, meaning no semen will be
present on the victims body, and genetic evidence from semen cannot, thus, be extracted.
In such cases all semen tests will fail. Alternative genetic evidence linking the perpetrator to
the victim could be collected, including saliva samples from places where the assailant has
licked the victim or cells from the perpetrator's penis that have been left behind in the
victim's vagina.
QUESTIONS FOR DNA APPLICATION
1) What type of technique shall be used for DNA testing?
2) Where shall the DNA testing be conducted?
3) What are your proofs that it is an accredited institution/agency to conduct DNA
testing?
4) Who will conduct?
5) Trainings attended?