Title: Forensic science and DNA finger printing

Aim: To determine how forensic science is being used and the methods that are applied in DNA
identification
Introduction
Forensic science is the scientific method of gathering and examining information about the past
which is then used in a court of law. Fingerprinting is the discipline that has been an important
part of police and forensic investigations. Fingerprints represent unique patterns of the ridges on
the pads of the fingers. The ridges occur in the epidermis but extend into the dermis. Barring
changes related to scar formation, which can obliterate portions of a fingerprint, fingerprints
remain the same throughout an individuals life. The presumption is that each individual has their
own unique set of fingerprints. No two fingerprints have ever been found to be exactly identical
even between identical twins. Therefore, a fingerprint represents a specific, individual
characteristic of a particular person. Fingerprint examiners rely on various class characteristics as
well as individual characteristics of fingerprints in their examinations. An evidence fingerprint at
a crime scene can be matched to a known print in a database. A variety of methods are used to
collect and preserve evidence fingerprints. For forensic pathologists, fingerprint comparison can
be extremely useful in identifying an unknown corpse.
Blood and other bodily fluids can be used to identify a person. Forensic serologists can perform
tests to determine if a suspicious fluid or stain is saliva, semen or blood. Tests are available to
determine if the evidence, such as blood, is of human origin. Once it is identified as human, then
DNA testing can be attempted.
Overview of Presentation
Karyotyping is a test to examine chromosomes in a sample of cells, which can help identify
genetic problems as the cause of a disorder or disease (www.geneticseducation). The karyotyping
test is done to numerate the number of chromosomes and identifying the structural changes in
chromosomes. The method used to perform this test first the sample is placed into a special dish
or tube and allowed to grow in the laboratory. Cells are later taken from the new sample and
stained. The laboratory specialist uses a microscope to examine the size, shape, and number of
chromosomes in the cell sample. The stained sample is photographed to shows the arrangement
of the chromosomes. This given what is known as the karyotype.
Multiplex polymerase chain reaction PCR can be defined as the amplifying multiple sequences
in a single reaction (Markoulatos P, Siafakas N, Moncany M, 2002). This process requires that
primers lead to amplification of unique regions of DNA, both in individual pairs and in
combinations of many primers, under a single set of reaction conditions. Also, the methods must
be available for the analysis of each individual amplification product from the mixture of all the
products. Multiplex PCR is becoming a rapid and convenient screening assay in both the clinical

and the research laboratory. An efficient multiplex PCR usually employs strategic planning and
repeating trial to optimize reaction conditions.
Multiplex PCR assay is affected by the concentration of the primers, concentration of the PCR
buffer, balance between the magnesium chloride and deoxynucleotide concentrations, cycling
temperatures, and amount of template DNA and Taq DNA polymerase. Magnesium chloride
concentration needs only to be proportional to the amount of dNTP. The list of various factors
that can influence the reaction is by no means complete. The sensitivity and specificity must be
thoroughly evaluated using standardized purified nucleic acids. Multiplex PCR reaction can be
divided single template PCR reaction and multiple template PCR reaction. For the single
template PCR used a single template which can be a genomic DNA along with several pairs of
forward and reverse primers to amplify specific regions within a template. While the multiple
templates PCR uses multiple templates and several primer sets in the same reaction tube.
Presence of multiple primers may lead to cross hybridization with each other and the possibility
of miss-priming with other templates (www.premierbiosoft.com).
Some of the advantages of using multiplex PCR are Internal Controls Potential by eliminating
false negatives and false positives due to contamination. False negatives are often revealed in
multiplex assays because each amplicon provides an internal control for the other amplified
fragments. One next advantage is efficiency as the expense of reagents and preparation time is
less in multiplex PCR than in systems where several tubes of PCRs are used. Multiplex also
indication of template quality as the quality of the template may be determined more effectively
in multiplex than in a simple PCR reaction. It is also true for the indication of Template Quantity
to quantitate templates accurately by multiplex PCR, the amount of reference template, the
number of reaction cycles, and the minimum inhibition of the theoretical doubling of product for
each cycle must be accounted.
Chromosomal abnormalities are disorders that affect the entire chromosomes, where large
segments of them are missing, duplicated, or otherwise altered. Some of these abnormalities
include Downs Syndrome, Turner syndrome and Williams syndrome. Williamss syndrome is a
rare genetic disorder that affects a child's growth, physical appearance, and cognitive
development. People who have Williams syndrome are missing genetic material from
chromosome 7, including the gene elastin. While, Turner syndrome is caused by a missing or
incomplete X chromosome. Also, Down syndrome is a developmental disorder caused by an
extra copy of chromosome 21 (learn.genetics.utah.edu).
Short Tandem Repeats are DNA regions with short repeat units (usually 2-6 bp in length).
They are usually found surrounding the chromosomal centromere .STRs is especially suitable for
human identification. STR is important DNA markers because they are easily amplified by PCR
without the problem of differential amplification. An individual inherits one copy of an STR
from each parent. The smaller size of STR alleles makes STR markers better candidates for use
in forensic applications, in which degraded DNA is common. PCR amplification of degraded

DNA samples can be better accomplished with smaller target product sizes. Because of their
smaller size, STR alleles can also be separated from other chromosomal locations more easily to
ensure closely linked loci are not chosen. Because of these characteristics, STRs with higher
power of discrimination are chosen for human identification in forensic cases on a regular basis.
It is used to identify victim, perpetrator, missing persons, and others
(www.forensicdnacenter.com).
QF-PCR is a laboratory technique used to copy small sections of DNA in order to precisely
quantify the amount of DNA present (. it work by coping small portion of the DNA ; typically 35 sections from chromosomes 13, 18, 21 and each sex chromosome. Primers used is labelled
with a different fluorescent tags and are different sizes. This enable researcher to determine
which parts of the chromosomes are represented. The amount of DNA in the sample can be
measured by QF PCR analysis includes amplification, detection and analysis of chromosomespecific DNA sequences known as genetic markers or small tandem repeats. Fluorescently
labelled marker specific primers are used for PCR amplification of individual markers and the
copy number of each marker is indicative of the copy number of the chromosome. The relative
copy number of each allele of STRs is determined by calculating the ratio of the peak areas or
peak heights detected for each marker. A normal diploid sample has the contribution of two of
each of the investigated chromosomes (Colman W & Tsongalis G, 2006). A 1:1 ratio of the two
chromosomal alleles is detected as two peaks, when the marker is heterozygous and as one peak
when the marker is homozygous.
The inability of STR marker analysis to distinguish subjects who are homozygous is a major
problem when testing for sex chromosome abnormalities. Has STRs specific for chromosome X
is used, some samples from normal females may show homozygous QF PCR patterns. These will
appear indistinguishable from those produced by samples with a single X, as in Turner
syndrome. Incorporating additional X-chromosome STR markers into the analysis will reduce
but not eliminate the likelihood of homozygosity. Some used of the QF-PCR are detection of
aneuploidies in antenatal amniocentesis, prenatal diagnosis and detect trisomies.
Parental testing is the use of genetic fingerprinting to determine whether two
individuals have a biological parentchild relationship (William J. Tilstone, Kathleen
A. Savage, Leigh A. Clark, 2006). A paternity test establishes genetic proof whether
a man is the biological father of an individual, and a maternity test establishes
whether a woman is the biological mother of an individual. Though genetic testing is
the most reliable standard, older methods also exist, including ABO blood group
typing, analysis of various other proteins and enzymes, or using human leukocyte
antigen antigens. The current techniques for paternal testing are using polymerase
chain reaction and restriction fragment length polymorphism (RFLP).
Paternity testing can also be performed while the woman is pregnant. First a
sample of a cheek cell is collected from both the child and the alleged father so that
DNA analysis can be done .after collection a pure DNA is isolated from the sample

by removing all the proteins and other things that can be found within a cell. Next,
the laboratory examines specific loci of each individual DNA sample. Two reading
will appear as each individual have 2 copies of each chromosome. DNA loci are
compared after testing is complete. The reading for the child will match to mother
and father if they are biological related. If the numbers match, a paternity index will
be determined. The paternity index is a calculation of how frequently that match
occurs in a specific race population, the likelihood that the tested man is the
biological father based on those loci. One locus is not enough to conclusively
determine paternity, so most labs test multiple loci. Each locus has its own paternity
index. If all the loci match, the paternity indices for each are combined, and a
probability of paternity is calculated. The probability of paternity is the final
percentage calculated.
References
Forensic Specialties Accreditation Board. http://www.thefsab.org retrieved 15/4/2015

Colman W & Tsongalis G, 2006 (Molecular Diagnostics: For the Clinical Laboratorian) humana
press.
William J. Tilstone, Kathleen A. Savage, Leigh A. Clark, 2006( Fetal Medicine: Basic

Science and Clinical Practice). Harcourt Brace and company.

Short Tandem Repeats STRs www.forensicdnacenter.com retrieved 14\4\2015

What is a chromosome learn.genetics.utah.edu) Retrieved 14\4\2015

Multiplex PCR www.premierbiosoft.com retrieved 14\4\2015
.