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TOPIC

DNA TYPING
INTRODUCTION
“ DNA typing is a procedure where in DNA extracted from a biological
sample obtained from an individual is analyzed. The DNA is processed
to generate a pattern for each person that is generally termed as a '
DNA profle'. This profle is uniue for each person excepting that
derived from identical twins.”
DNA testing is a relatively recent technological advance in the
feld of forensic biology The technology has a re!ar"able
#o$er to discern genetic di%erences& #retty $ell to the #oint
of individ'ali(ation Co'#led $ith the develo#!ent of
increasingly sensitive !ethods

)I*TORY O+ +OR,N*IC DNA ANA-Y*I*
DNA typing was introduced into forensic science in the mid !"#$s%
arising from discoveries made in biomedical research. Ray .hite% an
American geneticist% identifed regions of DNA that did not code for
proteins but were highly variable between individuals.
DNA fnger #rinting or DNA ty#ing /DNA #rofling0 $as describe
frst in 1234 by ,nglish geneticist Alec 5e%reys fo'nd that
certain region of DNA contained DNA se6'ence that $ere
re#eated over and over against ne7t to each other. &e also
discovered that the number of repeated section present in a sample
could di'er from individual to individual. 8y develo#ing a techni6'e
to e7a!ine the length variation of these DNA re#eat
se6'ences& Dr 5e%reys created the ability to #erfor! h'!an
identity tests
&uman identity test using DNA typing method has been widespread.
The past !( years have been seen tremendous growth in the use of
DNA evidence in crime scene investigation as well as paternity testing.
Today over !$$ public forensic laboratories and several dozen private
paternity testing laboratory conduct !$$ of !$$$ of DNA tests annually
in the )nited *tates.
9NTR
1
+The DNA repeat regions became ,nown as -NT.s% which stands for
variable number of tandem repeats./ 0ortions of the DNA contain
seuences of bases that are repeated numerous times. 1or example% a
base seuence such as AT2T may be repeated with in a stand of DNA
such as3
2A2AT2TAT2TAT2TAT2TAT2TA
T2TAT2TAT2TAT2TAT2TATT42
or
2A2AT2TAT2TAT2TAT2TAT2TA
T2TAT2TAT2TAT2TATT42
The origins of these tandem repeats are un,nown. VNTR loci are very
similar between closely related humans, but so variable that unrelated individuals are
extremely unlikely to have the same VNTRs. 5y some estimates% ""." percent
of the genetic code is the same in all humans. 6nly one7tenth of a
percent of DNA di'ers from one human to the next% To identify
individuals% DNA tests focus on a few loci where there is variation
among individuals. These loci are called #oly!or#his!s because
the genetic code can ta,e di'erent forms in di'erent individuals. 8ach
possible form is called an allele. 1orensic DNA tests have examined two
types of polymorphisms.
• *e6'ence #oly!or#his!s vary only in the seuence of the
genetic code.
• -ength #oly!or#his!s contain repeating seuences of
genetic code. The number of repetitions may vary from person to
person% ma,ing the section longer in some people and shorter in
others.
SOURCES OF DNA
DNA occurs in all living cells o our bodies, with the exce!tion o most red blood cells.
"owever, because our blood also contains white blood cells, DNA can be obtained rom
• #lood sam!les.
• $aliva.
• "air.
• $emen.
)o$ do yo' create a DNA #rofle/ DNA Ty#ing0:
%
*T,P* IN DNA *A;P-, PROC,**ING
8IO-OGY
1 The DNA is frst e7tracted fro! its biological so'rce
!aterial and then !eas'red to eval'ate the 6'antity of
DNA recovered After isolating the DNA fro! its cells&
s#ecifc
< regions are co#ied $ith a techni6'e "no$n as the
#oly!erase chain reactionPCR #rod'ces !illions of
co#ies for each starting DNA !olec'le and th's #er!its
very !in'te a!o'nts of DNA to be e7a!ined ;'lti#le
*TR regions can be e7a!ined si!'ltaneo'sly to increase
the infor!ativeness of the DNA test
T,C)NO-OGY
The resulting 02. products are then separated and detected in order to
char7acterize the *T. region being examined.
1 The se#aration !ethods 'sed today incl'de slab gel and
ca#illary electro#horesis /C,0
< .+l'orescence detection methods have greatly aided the
sensitivity and ease of measuring the number of repeats in a
DNA seuence is determined% a process ,nown as sa!#le
genoty#ing
Co!#arison=
The resulting DNA profle for a sample% which is a combination of
individual *T. genotypes% is compared to other samples. 9n the case of
a forensic investi7gation% these other samples would include ,nown
reference samples such as the victim or a suspect that are compared
to the crime scene evidence. :ith paternity investigations% the child;s
genotype would be compared to his or her mother;s and the father<s=
under investigation. 9f there is no match between the uestioned
sample and the ,nown sample% then the samples maybe considered to
have originated from di'erent sources. The term for failure to match
between two DNA profles is >exclusion.;
&
from3http?www. 1oresci.org? html
+OR,N*IC DNA TYPING T,C)NI>U,*
2urrently% There are four main types of forensic DNA testing3
• .1@0 <.estriction 1ragment @ength 0olymorphism=.
• 02. <0olymorphism 2hain .eaction=.
• *T. <*hort Tandem .epeat=.
• mtDNA <Aitochondrial DNA Analysis=.
4enerally .1@0 analysis reuires large amounts of DNA and the DNA
must be un7degraded. 02. testing often reuires less DNA than .1@0
testing and the DNA may be partially degraded. &owever% PCR still
has sa!#le si(e and degradation li!itations. PCR tests are
e7tre!ely sensitive to conta!ination at the cri!e scene and
$ithin the laboratory. *T. is one of the newer and more Bexible
DNA techniues. 9t has the advantage of being able to analyze
degraded and bro,en pieces of DNA. The fourth test type% mtDNA% is
used to examine samples which can not be analyzed by other methods
by loo,ing at the DNA in a cell's mitochondrion <as opposed the DNA in
the cell nucleus=.
 RFLP ANALYSIS
R+-P is not often 'sed in c'rrent cri!e scene analysis It
re6'ires relatively large a!o'nts of sa!#le Additionally& a
'
sa!#le that is old or degraded by environ!ental factors does
not $or" $ell $ith this #rocess
.1@0 as a techniue analyzes variable lengths of DNA
frag!ents that res'lt fro! digesting a DNA sa!#le
$ith s#ecifc en(y!es This en(y!e& restriction
endon'clease& brea"s DNA at a s#ecifc se6'ence
#attern "no$n as a restriction endon'clease
recognition site
The result is usually a set of -NT.'s. The presence or absence of
certain recognition sites in a DNA sample generates di'erent lengths of
DNA fragments% which may be separated by gel electrophoresis.
They are then hybridi(ed with DNA probes that bind to
complementary DNA seuences in the sample. 1or .1@0 analysis to be
reliable% all steps of the analysis must be carefully controlled. 9n
addition% databases must be suCciently large and representative of the
relevant population.
 PCR ANALYSIS
02. analysis is a techniue that allows technicians to create millions of
precise DNA replications from a single sample of DNA. 9n fact% DNA
amplifcation alongside 02. can let forensic scientists perform. In
contrast to so!e other DNA analysis techni6'es& PCR analysis
has the advantage of analy(ing !in'sc'le sa!#le si(es& even if
they are degraded altho'gh they !'st not be conta!inated
$ith DNA fro! other so'rces d'ring the collection& storage and
trans#ort of the sa!#le
 STR ANALYSIS
$hort tandem re!eats (or $TRs) are regions o noncoding DNA that contain re!eats o the
same nucleotide se*uence.+or exam!le,
,ATA,ATA,ATA,ATA,ATA,ATA is a $TR where the n'cleotide
se6'ence ,ATA is re!eated six times.
*T. are found at di'erent places or genetic loci in a person;s DNA.*T.
analysis wor,s to examine individual areas in DNA. The di%erences
fro! the collective areas of one #erson to another can allo$
for disting'ishing bet$een individ'als
In cri!inal investigations& there are thirteen regions that are
analy(ed and co!#ared to establish #rofles
In fact& DNA databases 'sed at the govern!ent level involve
the se6'ence of these thirteen regions
The chances of t$o #eo#le having the e7act sa!e thirteen
regions is virt'ally i!#ossible ? li"ely one in a billion
-
 ;ITOC)ONDRIA- DNA ANA-Y*I*
Aitochondrial DNA analysis works well on sam!les that are unable to be analysed
through R+./ or $TR analysis. There are two kinds o DNA in the cell 0 mitochondrial
DNA and nuclear DNA. 1ith other ty!es o analysis, nuclear DNA is removed rom the
sam!le but with mitochondrial DNA analysis, DNA is removed rom the cell2s
mitochondria. $ometimes, a sam!le can be old and will no longer have nuclear material
in the cell, which !oses a !roblem or the other ty!es o DNA analysis. 1ith
mitochondrial DNA analysis, however, mitochondrial DNA can be removed, thus having
im!ortant ramiications or cases that were not solved over many years. This means that
mitochondrial DNA analysis can be very valuable in investigations or a missing !erson.
Mitochondrial DNA will be the sae !ro a woan to her da"#hter beca"se it is
$assed on !ro the e## cell%
There are other ty!es o analysis techni*ue also used but these are some o the main
traditional and current methods used to analyse DNA. No doubt, new techni*ues will be
develo!ed that will be even more ra!id, successul and cost0eective.
APP-ICATION O+ DNA TYPING
DNA typing has broad applications. This procedure is used in countries
li,e )*A% Australia% 4ermany% 2anada and many others to verify the
li,ely owner of a piece of tissue such as blood% hair% bits of Besh% bone%
teeth% semen or any other part of the body.
2learly besides disputed parentage cases% DNA typing also aids
• in criminal investigations
• in identifcation of mass disaster victims
• in verifying the identity of human cancer cell lines
• in determining whether a biological material is of human origin
• in studying the genetic ancestry of human populations.

3
G,N, T),RAPY
INTRODUCTION
The prospect of human gene therapy was frst realized in !"D! when
the frst recombinant DNA experiments were planned. 4ene
therapy is currently an experimental discipline. AaEor advances
in reco!binant DNA technology have occurred over the last
F$ years so that now gene therapy is becoming a reality.
4enes are the blueprint for our bodies% providing information for the
cells to produce proteins and enzymes to control our growth%
development and health. A genetic mutation means that a gene
contains a variation or >spelling mista,e; that disrupts the gene
message. *ometimes% the whole or part of the gene is missing
<deleted=. These changes can ma,e the gene faulty. A mutation
can occur spontaneously or may be inherited.. The defnition of
gene therapy3
 @Techni6'e $here the genes ca'sing a defect are
the!selves s'bstit'ted by correct genes in the
#atient to c're a disease@
TYP,* O+ G,N, T),RAPY
The underlying principle of gene therapy is the transfer of genetic
material to specifc cells of a patient in an e'ort to initiate a
biological res#onse to fght or eliminate a disease. 9n theory% it
is possible to insert genes into either3
 *O;ATIC C,-- G,N, T),RAPY
As the na!e s'ggests& in so!atic gene thera#y& the
thera#e'tic genes are transferred into the so!atic cells /non
se7?cells0& or body& of a #atient Any !odifcations and e%ects
$ill be restricted to the individ'al #atient only& and $ill not be
inherited by the #atientAs o%s#ring or later generations
 G,R;-IN, C,-- G,N, T),RAPY
In ger!line gene thera#y& ger! cells /s#er! or eggs0 are
!odifed by the introd'ction of f'nctional genes& $hich are
integrated into their geno!es Ger! cells $ill co!bine to for!
a (ygote $hich $ill divide to #rod'ce all the other cells in an
organis! and therefore if a ger! cell is genetically !odifed
4
then all the cells in the organis! $ill contain the !odifed
gene This $o'ld allo$ the thera#y to be heritable and #assed
on to later generations
8A*IC *T,P*
4ene therapy is an experimental techniue that uses genes to treat or
prevent disease. 9n the future% this techniue may allow doctors to
treat a disorder by inserting a gene into a patient;s cells instead of
using drugs or surgery. The most complex phase in gene
therapy is the development of mechanisms to deliver the
therapeutic genes to the target organ in an accurate,
controlled, and efective way. .esearcher are testing several
approaches to gene therapy.
The basic steps of gene therapy include3
• The faulty gene that causes a specifc condition must be
identifed
• The location of the a%ected cells in the bodyBs tiss'es or
organs !'st be #in#ointed
• A $or"ing version of the gene !'st be available
• The wor,ing version of the gene has to be delivered to the cell.
• .eplacing a mutated gene that causes disease with a healthy
copy of the gene.
• 9nactivating% or +,noc,ing out%/ a mutated gene that is
functioning improperly.
• 9ntroducing a new gene into the body to help fght a disease.
Although gene therapy is a promising treatment option for a number of
diseases. About ',555 diseases have been traced to gene disorders. 6urrent and
!ossible candidates or gene thera!y include7
,CA;P-,
6ancer, A8D$, cystic ibrosis, /arkinson9s and Al:heimer9s diseases <including
inherited disorders% some types of cancer% and certain viral
infections=%the techniue remains ris,y and is still under study to ma,e
sure that it will be safe and e'ective. 4ene therapy is currently only
being tested for the treatment of diseases that have no other cures.
The irst successul gene thera!y on humans was !erormed in 1;;5 by researchers at
the National 8nstitutes o "ealth. The thera!y treated a our0year0old child or adenosine
deaminase (ADA) deiciency, a rare genetic disease in which children are born with
severe immunodeiciency lacks an im!ortant en:yme o their immune system and are
!rone to re!eated serious inections.
T,C)NO-OGICA- A*P,CT* O+ G,N, T),RAPY
.
<
Gene thera#y is designed to introd'ce genetic !aterial into
cells to co!#ensate for abnor!al genes or to !a"e a
benefcial #rotein. 9f a mutated gene causes a necessary protein
to be faulty or missing% gene therapy may be able to introduce a
normal copy of the gene to restore the function of the protein.
4ene therapy utilizes the delivery of DNA into cells% which can be
accomplished by a number of methods.
The two maEor classes of methods are those that use recombinant
viruses <sometimes called biological nanoparticles or viral vectors= and
those that use na,ed DNA or DNA complexes <non7viral methods=.
NON? 9IRA- ;,T)OD*
Non7viral methods can present certain advantages over viral methods%
such as large scale #rod'ction and lo$ host i!!'nogenicity.
There are several methods for non7viral gene therapy% including
0&G*92A@A8T&6D
• The inEection of na,ed DNA%
• 8lectroporation% (is a signiicant increase in the electrical conductivity and
!ermeability o the cell !lasma membrane caused by an externally a!!lied
electrical ield. 8t is usually used in molecular biology as a way o introducing
some substance into a cell)
• The use of gene gun.
• $ono!oration uses ultrasonic re*uencies to deliver DNA into cells. The !rocess o
acoustic cavitations is thought to disru!t the cell membrane and allow DNA to
move into cells.
2&8A92A@ A8T&6D
• The use of oligonucleotides (are short, single0stranded DNA or RNA
molecules can be manuactured with any user0s!eciied se*uence, and so are vital
or artiicial gene synthesis=.
• Li$o$le&es ' A li!osome is an artiicially0!re!ared s!herical vesicle com!osed o a
li!id bilayer. The li!osome can be used as a vehicle or gene transer. Pol($le&es'
6om!lexes o !olymers with DNA are called !oly!lexes.
9IRA- ;,T)OD
VECTORS ! "E!E T#ER$%&
;
A )ector is si$l( a *trans$orter* !or the #enetic aterial that allows it to enter the
tar#et cell and+ de$endin# on the )ector t($e+ can ca"se new #enes to be inte#rated
into the host cell #enoe%
6ertain viruses are used as vectors because they can deliver the new gene by inecting
the cell. virus as a vehicle to deliver the thera!eutic DNA. =ne !romising techni*ue is to
!ut the working gene inside a harmless virus, which has had most o its own genes
removed > it has been ?deactivated9. The viruses are modiied so they can9t cause disease
when used in !eo!le. $cientists must ind better ways to deliver genes and target them to
!articular cells. A number o viruses have been used or human gene thera!y,
2haracteristics of -iruses That &ave 5een )sed to 4enerate -iral
-ector
)O. DO,* G,N, T),RAPY .ORD
Doctors and molecular biologists realized that viruses li,e this could be
used as vehicles to carry >good; genes into a human cell scientists
retain the outer viral coat% but modify the inner genetic material. They
remove the harmful genes and replace them with therapeutic ones.
Now the virus is pathogenically disabled <it is no longer harmful to the
cell it infects= and incapable of reproducing itself. )o$ever& it retains
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its ca#ability to transfer its genetic !aterial to the cells for
$hich its o'ter coat $as designed The transfer of genetic
!aterial by $ay of a viral vector is called transduction
8H -9-6 A8T&6D3 cells are genetically7altered outside of the body and
then reintroduced.
Adapted
from3http?www.elsine.org?idDI html
9N -9-6 A8T&6D3 cells are genetically7altered inside the body after a
genetically7altered vector containing therapeutic DNA is inEected into
the body.
11
Adapted from3http?www.elsine.org?idDI html
T), RI*D* O+ G,N, T),RAPY
*ome of these ris,s may include3
• The i!!'ne syste! !ay res#ond to the $or"ing gene co#y
that has been inserted by ca'sing inEa!!ation
• The $or"ing gene !ight be slotted into the $rong s#ot
• The $or"ing gene !ight #rod'ce too !'ch of the !issing
en(y!e or #rotein& ca'sing other health #roble!s.
• Other genes !ay be accidentally delivered to the cell
• The deactivated vir's !ight target other cells as $ell as the
intended cells
• The deactivated vir's !ay be contagio's
CONC-U*ION
DNA Typing is modern techniue play an important role in modern
scenario The technology has a remar,able power to discern genetic
di'erences% pretty well to the point of individualization.4ene therapy is
consider as modern medicine . 9t is an experimental discipline help in
cure diseases.
R,+R,NC,*
• 2ann% .. @. J :ilson% A. 2. 4enetics !$K% I""LD!! <!"#M=.
1%
• David &. Naye and 4eorge 1. *ensabaugh% Or.% .eference 4uide
on DNA 8vidence% at K#(% K"! in .eference Aanual on *cientiic
8vidence <Fd 8d. :est 4roup% F$$$=.
• 1riedmann% T.P .oblin% .. <!"DF=. Q4ene Therapy for &uman
4enetic DiseaseRQ. *cience !D( <K$F(=3 "K"S"((.
• Oe'reys A.O.% :ilson -.% Thein *.:. <!"#K=. Q&ypervariable
'minisatellite' regions in human DNAQ. Nature M!K3 IDSDM.
• Oames A. :ilson. 9t's Time for 4ene Therapy to 4et
Disruptive.&uman 4ene Therapy, F$!FP FM <!=.
• :ong% T. et al. Ann. hum. 4enet. (!% FI"LF## <!"#D=.
1&