NAME – ADHIRAJ SINGH CHAUHAN

CLASS – B.A.LLB Div. ‘B’ YEAR ‘IV’

PRN – 14010125102

SUBJECT – FORENSIC SCIENCE

Q.1. Explain how PCR is used to amplify DNA molecules.

Ans. Sometimes called "molecular photocopying," the polymerase chain reaction (PCR) is a
fast and inexpensive technique used to "amplify" - copy - small segments of DNA. Because
significant amounts of a sample of DNA are necessary for molecular and genetic analyses,
studies of isolated pieces of DNA are nearly impossible without PCR amplification.

Once amplified, the DNA produced by PCR can be used in many different laboratory
procedures. For example, most mapping techniques in the Human Genome Project (HGP)
relied on PCR. PCR is also valuable in a number of laboratory and clinical techniques,
including DNA fingerprinting, detection of bacteria or viruses (particularly AIDS), and
diagnosis of genetic disorders.

To amplify a segment of DNA using PCR, the sample is first heated so the DNA denatures,
or separates into two pieces of single-stranded DNA. Next, an enzyme called "Taq
polymerase" synthesizes - builds - two new strands of DNA, using the original strands as
templates. This process results in the duplication of the original DNA, with each of the new
molecules containing one old and one new strand of DNA. Then each of these strands can be
used to create two new copies, and so on, and so on. The cycle of denaturing and synthesizing
new DNA is repeated as many as 30 or 40 times, leading to more than one billion exact
copies of the original DNA segment.

The entire cycling process of PCR is automated and can be completed in just a few hours. It
is directed by a machine called a thermo cycler, which is programmed to alter the temperature
of the reaction every few minutes to allow DNA denaturing and synthesis.

Q.2. Explain how STRs are used in DNA profiling.

Ans. A Short Tandem Repeat (STR) analysis is one of the most useful methods in molecular
biology which is used to compare specific loci on DNA from two or more samples. A short
tandem repeat is a microsatellite, consisting of a unit of two to thirteen nucleotides repeated
hundreds of times in a row on the DNA strand. STR analysis measures the exact number of
repeating units. This method differs from restriction fragment length polymorphism
analysis (RFLP) since STR analysis does not cut the DNA with restriction enzymes. Instead,
probes are attached to desired regions on the DNA, and a polymerase chain reaction (PCR) is
employed to discover the lengths of the short tandem repeats. The most common type of
DNA profiling today for criminal cases and other types of forensic uses is called "STR"
(short tandem repeat) analysis.

STR analysis is a tool in forensic analysis that evaluates specific STR regions found
on nuclear DNA. The variable (polymorphic) nature of the STR regions that are analysed for
forensic testing intensifies the discrimination between one DNA profile and another. Forensic
science takes advantage of the population's variability in STR lengths, enabling scientists to
distinguish one DNA sample from another. The system of DNA profiling used today is based
on PCR and uses simple sequences or short tandem repeats (STR). This method uses highly
polymorphic regions that have short repeated sequences of DNA (the most common is 4
bases repeated, but there are other lengths in use, including 3 and 5 bases). Because unrelated
people almost certainly have different numbers of repeat units, STRs can be used to
discriminate between unrelated individuals. These STR loci (locations on a chromosome) are
targeted with sequence-specific primers and amplified using PCR. The DNA fragments that
result are then separated and detected using electrophoresis. There are two common methods
of separation and detection, capillary electrophoresis (CE) and gel electrophoresis.

Each STR is polymorphic, but the number of alleles is very small. Typically each STR allele
will be shared by around 5 - 20% of individuals. The power of STR analysis comes from
looking at multiple STR loci simultaneously. The pattern of alleles can identify an individual
quite accurately. Thus STR analysis provides an excellent identification tool. The more STR
regions that are tested in an individual the more discriminating the test becomes.

In practice, the risk of contaminated-matching is much greater than matching a distant

relative, such as contamination of a sample from nearby objects, or from left-over cells
transferred from a prior test. The risk is greater for matching the most common person in the
samples: Everything collected from, or in contact with, a victim is a major source of
contamination for any other samples brought into a lab. For that reason, multiple control-
samples are typically tested in order to ensure that they stayed clean, when prepared during
the same period as the actual test samples. Unexpected matches (or variations) in several
control-samples indicates a high probability of contamination for the actual test samples. In a
relationship test, the full DNA profiles should differ (except for twins), to prove that a person
was not actually matched as being related to their own DNA in another sample.

Q.3. Use DNA profiling results to match a suspect to a crime scene.

Ans. The DNA molecule, present in every cell in our bodies, is a chain of molecules, sugars,
and phosphates. The order in which these components arrange themselves in the chain is,
statistically speaking, unique to every individual (except for identical twins). The DNA
molecule is the same in every cell, and remains constant (with slight, rare variations) during
an individual’s life. Scientists can harvest a person’s DNA molecule from any cell, be it
tissue, blood, sweat, semen, bone, saliva, and so on. Once the DNA ladder is analyzed, it can
be compared to another molecule’s DNA. Forensic scientists perform such comparisons in
order to:

 Link a suspect to a crime or exonerate someone, where biological evidence has been
left at the crime, by comparing the suspect’s DNA to that left at the scene

 Determine parentage, for purposes of paternity, immigration, and other cases, and

 Identify human remains, when visual or dental determinations are not possible or
conclusive.

DNA identification is extraordinarily reliable, though it can be challenged, as explained

below.

In the lab, scientists extract an amount of DNA from each biological sample, which must be
sufficiently large—a quarter-size blood stain, or a dime-sized semen stain. The DNA
molecule itself must be intact, not in pieces that are too small to analyze. Then, each sample’s
band pattern (its sequence) is compared to the other. If the bands match, scientists consult a
national database of band patterns, and determine the statistical chances that the match is a
coincidence. In court, you’ll hear the scientist testify that the odds of the match being
coincidental are, for instance, many millions-to-one, leaving the jury with the evidence it
needs to accept the match as correct. (Another type of analysis can be performed on a smaller
sample, such as a single hair, with results that may be less convincing.)

The process of collecting, testing, and interpreting DNA samples is not foolproof. Samples
are collected literally “in the field,” not in pristine laboratories. Many factors can influence
the reliability of the results. They include:

 Degradation due to age, environment, contamination. The sample collected at a

crime scene or elsewhere may be old, dried-up, soggy, or sullied by grease, dirt, or
other contaminants. These are factors that may break the DNA into small fragments,
reducing the clarity of the bands or their sizes.

 Variations in interpretation of the bands due to human subjectivity. In the end, a

human compares the samples’ bands and declares a match—or not. When a sample
has been degraded, the analyst lacks a crisp print, which makes it possible to
mistakenly declare a match.

 Lack of consensus as to the basis for statistical comparisons. A match derives its
persuasive force from the analyst’s ability to tell the judge or jury that the odds of a
coincidental match are astronomical. Yet within the scientific community, a debate
exists as to the basis for these statistical pronouncements.

 Qualifications of the lab techs. The tests will only be as good as those doing the
work—did the lab technicians correctly follow accepted procedures?

 Qualifications of the people doing the analysis. To establish the reliability of any
test, the proponent should offer the testimony of a qualified molecular biologist and a
 population geneticist. Ideally, these witnesses should be from the academic
community, not from the private lab that did the work, because those working in the
private sector arguably have a financial interest in the acceptance of their work.

Q.4. Use DNA profiling results to match a suspect to a crime scene.

Ans. DNA profiling and matching of physical data, such as fingerprints, are used in solving
all crime types ranging from housebreaking and car crime to assaults, murder and rape. The
forensic scientists will look for suitable samples at a crime scene, examining such items as
weapons, clothing, hair or anything else from which they can obtain body cells for DNA
profiling, or fingerprints or "marks" for use in fingerprint matching.

The past decade has seen great advances in a powerful criminal justice tool: deoxyribonucleic
acid, or DNA. DNA can be used to identify criminals with incredible accuracy when
biological evidence exists. By the same token, DNA can be used to clear suspects and
exonerate persons mistakenly accused or convicted of crimes. In all, DNA technology is
increasingly vital to ensuring accuracy and fairness in the criminal justice system.

News stories extolling the successful use of DNA to solve crimes abound. For example, in
1999, New York authorities linked a man through DNA evidence to at least 22 sexual
assaults and robberies that had terrorized the city. In 2002, authorities in Philadelphia,
Pennsylvania, and Fort Collins, Colorado, used DNA evidence to link and solve a series of
crimes (rapes and a murder) perpetrated by the same individual. In the 2001 “Green River”
killings, DNA evidence provided a major breakthrough in a series of crimes that had
remained unsolved for years despite a large law enforcement task force and a $15 million
investigation.

DNA is generally used to solve crimes in one of two ways. In cases where a suspect is
identified, a sample of that person’s DNA can be compared to evidence from the crime scene.
The results of this comparison may help establish whether the suspect committed the crime.
In cases where a suspect has not yet been identified, biological evidence from the crime scene
can be analyzed and compared to offender profiles in DNA databases to help identify the
perpetrator. Crime scene evidence can also be linked to other crime scenes through the use of
DNA databases.

For example, assume that a man was convicted of sexual assault. At the time of his
conviction, he was required to provide a sample of his DNA, and the resulting DNA profile
was entered into a DNA database. Several years later, another sexual assault was committed.
A Sexual Assault Nurse Examiner worked with the victim and was able to obtain biological
evidence from the rape. This evidence was analyzed, the resulting profile was run against a
DNA database, and a match was made to the man’s DNA profile. He was apprehended, tried,
and sentenced for his second crime. In this hypothetical case, he was also prevented from
committing other crimes during the period of his incarceration.
DNA evidence is generally linked to DNA offender profiles through DNA databases. In the
late 1980s, the federal government laid the groundwork for a system of national, state, and
local DNA databases for the storage and exchange of DNA profiles. This system, called the
Combined DNA Index System (CODIS), maintains DNA profiles obtained under the federal,
state, and local systems in a set of databases that are available to law enforcement agencies
across the country for law enforcement purposes. CODIS can compare crime scene evidence
to a database of DNA profiles obtained from convicted offenders. CODIS can also link DNA
evidence obtained from different crime scenes, thereby identifying serial criminals.

In order to take advantage of the investigative potential of CODIS, in the late 1980s and early
1990s, states began passing laws requiring offenders convicted of certain offenses to provide
DNA samples. Currently all 50 states and the federal government have laws requiring that
DNA samples be collected from some categories of offenders.

When used to its full potential, DNA evidence will help solve and may even prevent some of
the Nation’s most serious violent crimes. However, the current federal and state DNA
collection and analysis system needs improvement:

(1) In many instances, public crime labs are overwhelmed by backlogs of unanalyzed DNA
samples.

(2) In addition, these labs may be ill-equipped to handle the increasing influx of DNA
samples and evidence. The problems of backlogs and lack of up-to-date technology result in
significant delays in the administration of justice.

(3) More research is needed to develop faster methods for analyzing DNA evidence.

(4) Professionals working in the criminal justice system need additional training and
assistance in order to ensure the optimal use of DNA evidence to solve crimes and assist
victims.