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Dna Profiling Part 1

Dna Profiling Part 1

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10/17/2011

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DNA PROFILING & ITS

DNA PROFILING & ITS
APPLICATIONS
APPLICATIONS

DNA PROFILING
DNA PROFILING

Genetic
Genetic
Fingerprinting
Fingerprinting
(also
(also
called
called
DNA testing
DNA testing
,
,
DNA typing
DNA typing
, or
, or
DNA
DNA
profiling
profiling
) is a
) is a
technique used to
technique used to
distinguish beteen
distinguish beteen
indi!iduals o" the
indi!iduals o" the
sa#e s$ecies using
sa#e s$ecies using
onl% sa#$les o" their
onl% sa#$les o" their
DNA&
DNA&

'ho In!ented it(
'ho In!ented it(

)he $rocess o" DNA
)he $rocess o" DNA
"inger$rinting as
"inger$rinting as
in!ented b% Alec
in!ented b% Alec
*e""re%s at the
*e""re%s at the
+ni!ersit% o" Leicester
+ni!ersit% o" Leicester
in ,-./&
in ,-./&

0e as 1nighted in
0e as 1nighted in
,--2&
,--2&


)o understand DNA $ro"iling, %ou "irst ha!e to 1no that )o understand DNA $ro"iling, %ou "irst ha!e to 1no that
large $ortions o" an% single $erson3s DNA are the sa#e as large $ortions o" an% single $erson3s DNA are the sa#e as
e!er% other $erson3s& 4ecause e3re all hu#an beings, a e!er% other $erson3s& 4ecause e3re all hu#an beings, a
large chun1 o" our DNA is dedicated to our s$ecies5 large chun1 o" our DNA is dedicated to our s$ecies5
s$eci"ic traits 5 e ha!e "eet instead o" hoo!es, s1in instead s$eci"ic traits 5 e ha!e "eet instead o" hoo!es, s1in instead
o" scales, etc& 4ut other sections 5 or "rag#ents 5 o" hu#an o" scales, etc& 4ut other sections 5 or "rag#ents 5 o" hu#an
DNA are unique to the indi!idual& )hese "rag#ents are DNA are unique to the indi!idual& )hese "rag#ents are
called $ol%#or$hic because the% !ar% in sha$e "ro# called $ol%#or$hic because the% !ar% in sha$e "ro#
$erson to $erson& 6ssentiall%, DNA $ro"iling is the $rocess $erson to $erson& 6ssentiall%, DNA $ro"iling is the $rocess
o" se$arating an indi!idual3s unique, $ol%#or$hic, o" se$arating an indi!idual3s unique, $ol%#or$hic,
"rag#ents "ro# the co##on ones& "rag#ents "ro# the co##on ones&


Although to indi!iduals ill ha!e the !ast #a7orit% o" their Although to indi!iduals ill ha!e the !ast #a7orit% o" their
DNA sequence in co##on, DNA $ro"iling e8$loits highl% DNA sequence in co##on, DNA $ro"iling e8$loits highl%
!ariable re$eat sequences called !ariable re$eat sequences called VNTRs. VNTRs.

)hese loci are !ariable enough that to unrelated hu#ans are )hese loci are !ariable enough that to unrelated hu#ans are
unli1el% to ha!e the sa#e alleles& unli1el% to ha!e the sa#e alleles&
9 re$eats
. re$eats
AATG


6ach o" us has a unique DNA $ro"ile or "inger$rint& A
6ach o" us has a unique DNA $ro"ile or "inger$rint& A
technique called electro$horesis is used to obtain
technique called electro$horesis is used to obtain
DNA $ro"iles, rel%ing on sections o" our DNA that
DNA $ro"iles, rel%ing on sections o" our DNA that
are 1non as non5coding DNA : that does not code
are 1non as non5coding DNA : that does not code
"or a $rotein&
"or a $rotein&

For e8a#$le, %ou #a% ha!e a stretch o" DNA #ade
u$ o" the "olloing base sequence;

A)<))<)AA<A<A)GA<<GA)
A)<))<)AA<A<A)GA<<GA)
<A)G
<A)G
<
<
A)G<A)G<A)G<A)G<A)G<A)G<A)
A)G<A)G<A)G<A)G<A)G<A)G<A)
G<A)G<A)G<A)G))<<A)GA)AG<A
G<A)G<A)G<A)G))<<A)GA)AG<A
<A)
<A)


Pol%#erase chain reaction (P<R) is used "irst to
Pol%#erase chain reaction (P<R) is used "irst to
$roduce #an% co$ies o" the ten =)Rs be"ore the% are
$roduce #an% co$ies o" the ten =)Rs be"ore the% are
anal%>ed using electro$horesis& )he di""erent lengths
anal%>ed using electro$horesis& )he di""erent lengths
ill sho u$ as bands at di""erent s$ots on the
ill sho u$ as bands at di""erent s$ots on the
electro$horesis gel& )he banding $attern $roduced is
electro$horesis gel& )he banding $attern $roduced is
called a DNA $ro"ile or "inger$rint, and can be
called a DNA $ro"ile or "inger$rint, and can be
anal%>ed&
anal%>ed&

Ho c!n DNA "e #se$ to i$entify
Ho c!n DNA "e #se$ to i$entify
!n in$i%i$#!l&
!n in$i%i$#!l&

6!er% single cell in our bodies contains DNA, the
6!er% single cell in our bodies contains DNA, the
genetic #aterial that $rogra#s ho cells or1&
genetic #aterial that $rogra#s ho cells or1&
--&- $ercent o" hu#an DNA is the sa#e in
--&- $ercent o" hu#an DNA is the sa#e in
e!er%one, #eaning that onl% ?&, $ercent o" our
e!er%one, #eaning that onl% ?&, $ercent o" our
DNA is unique&
DNA is unique&

6ach hu#an cell contains three billion DNA base
6ach hu#an cell contains three billion DNA base
$airs& Our unique DNA, ?&, $ercent o" @ billion,
$airs& Our unique DNA, ?&, $ercent o" @ billion,
a#ounts to @ #illion base $airs& )his is #ore than
a#ounts to @ #illion base $airs& )his is #ore than
enough to $ro!ide $ro"iles that accuratel% identi"%
enough to $ro!ide $ro"iles that accuratel% identi"%
a $erson& )he onl% e8ce$tion is identical tins,
a $erson& )he onl% e8ce$tion is identical tins,
ho share ,?? $ercent identical DNA&
ho share ,?? $ercent identical DNA&


At a cri#e scene, DNA is e!er%here& It is $resent in
At a cri#e scene, DNA is e!er%here& It is $resent in
all 1inds o" e!idence collected at the scene, including
all 1inds o" e!idence collected at the scene, including
blood, hair, s1in, sali!a and se#en& =cientists can
blood, hair, s1in, sali!a and se#en& =cientists can
anal%>e the DNA in e!idence sa#$les to see i" it
anal%>e the DNA in e!idence sa#$les to see i" it
#atches a sus$ect3s DNA&
#atches a sus$ect3s DNA&

4iological #aterials used "or DNA
4iological #aterials used "or DNA
$ro"iling
$ro"iling

4lood 4lood

0air 0air

=ali!a =ali!a

=e#en =e#en

4od% tissue cells 4od% tissue cells

DNA sa#$les ha!e been DNA sa#$les ha!e been
obtained "ro# !aginal obtained "ro# !aginal
cells trans"erred to the cells trans"erred to the
outside o" a condo# outside o" a condo#
during intercourse& during intercourse&

=tages o" DNA Pro"iling
=tages o" DNA Pro"iling

St!ge ' St!ge '; ;
<ells are bro1en don <ells are bro1en don
to release DNA to release DNA
I" onl% a s#all a#ount o" I" onl% a s#all a#ount o"
DNA is a!ailable it can be DNA is a!ailable it can be
a#$li"ied using the a#$li"ied using the
$ol%#erase chain reaction $ol%#erase chain reaction
(P<R) (P<R)

=tages o" DNA Pro"iling
=tages o" DNA Pro"iling

Step () Step ()
)he DNA is cut into "rag#ents using )he DNA is cut into "rag#ents using restriction en*y+es restriction en*y+es& &
6ach restriction en>%#e cuts DNA at a s$eci"ic base 6ach restriction en>%#e cuts DNA at a s$eci"ic base
sequence& sequence&

=tages o" DNA Pro"iling
=tages o" DNA Pro"iling

)he sections o" DNA that are cut out are
)he sections o" DNA that are cut out are
called
called
restriction fr!g+ents.
restriction fr!g+ents.

)his %ields thousands o" restriction
)his %ields thousands o" restriction
"rag#ents o" all di""erent si>es because the
"rag#ents o" all di""erent si>es because the
base sequences being cut #a% be "ar a$art
base sequences being cut #a% be "ar a$art
(long "rag#ent) or close together (short
(long "rag#ent) or close together (short
"rag#ent)&
"rag#ent)&

=tages o" DNA Pro"iling
=tages o" DNA Pro"iling
St!ge ,)
St!ge ,)

Frag#ents are
Frag#ents are
se$arated on the basis
se$arated on the basis
o" si>e using a $rocess
o" si>e using a $rocess
called
called
gel
gel
electro$horesis&
electro$horesis&

DNA "rag#ents are
DNA "rag#ents are
in7ected into ells and
in7ected into ells and
an electric current is
an electric current is
a$$lied along the gel&
a$$lied along the gel&

=tages o" DNA Pro"iling
=tages o" DNA Pro"iling

DNA is negati!el%
DNA is negati!el%
charged so it is
charged so it is
attracted to the
attracted to the
$ositi!e end o" the gel&
$ositi!e end o" the gel&
)he shorter DNA
)he shorter DNA
"rag#ents #o!e "aster
"rag#ents #o!e "aster
than the longer
than the longer
"rag#ents&
"rag#ents&
DNA is se$arated on
DNA is se$arated on
basis o" si>e&
basis o" si>e&

=tages o" DNA Pro"iling
=tages o" DNA Pro"iling

A radioacti!e #aterial
A radioacti!e #aterial
is added hich
is added hich
co#bines ith the
co#bines ith the
DNA "rag#ents to
DNA "rag#ents to
$roduce a "luorescent
$roduce a "luorescent
i#age&
i#age&

A $hotogra$hic co$%
A $hotogra$hic co$%
o" the DNA bands is
o" the DNA bands is
obtained&
obtained&

=tages o" DNA Pro"iling
=tages o" DNA Pro"iling
St!ge -)
St!ge -)

)he $attern o" "rag#ent distribution is then
)he $attern o" "rag#ent distribution is then
anal%sed&
anal%sed&

+ses o" DNA Pro"iling
+ses o" DNA Pro"iling

DNA $ro"iling is used to DNA $ro"iling is used to
sol!e sol!e cri+es cri+es and and +e$ic!l +e$ic!l
pro"le+s pro"le+s

Parentage testing Parentage testing
(explored in more (explored in more
detail) detail)

Victim identification Victim identification
in mass disasters in mass disasters

Animal identification- Animal identification-
e.g. racehorses e.g. racehorses

Conservation biology Conservation biology
and evolutionary and evolutionary
studies studies


./THODOLOG0

DNA "inger$rinting begins b% e8tracting
DNA "ro# the cells in a sa#$le o" blood,
sali!a, se#en, or other a$$ro$riate "luid or
tissue&

./THODS

RFLP : ( Restriction Frag#ent
Length Pol%#or$his# 5
$ronounced as Ari"5li$A )

P<R : (Pol%#erase chain
reaction )

B5 chro#oso#e anal%sis

=)R : (short tande# re$eats)

a#$FLP

#itochondrial anal%sis

RFLP 1R/STRICTION FRAG./NT
L/NGTH POL0.ORPHIS. 2

)he "irst #ethods used "or DNA "inger$rinting
in!ol!ed restriction en>%#e digestion, "olloed
b% =outhern blot anal%sis&

Although $ol%#or$his#s can e8ist in the
restriction en>%#e clea!age sites, #ore
co##onl% the en>%#es and DNA $robes ere
used to anal%>e CN)R loci&


)he technique "or detecting RFLPs in!ol!es the
"rag#entation o" geno#ic DNA b% a restriction
en>%#e, hich can recogni>e and cut DNA
here!er a s$eci"ic short sequence occurs, in a
$rocess 1non as a restriction digest&

)he resulting DNA "rag#ents are then
se$arated b% length through a $rocess 1non as
agarose gel electro$horesis, D

trans"erred to a #e#brane !ia the =outhern blot
$rocedure
RFLP 3 T/CHNI45/

RFLP

0%bridi>ation o" the #e#brane to a labeled
DNA $robe then deter#ines the si>e o" the
"rag#ents hich are co#$le#entar% to the
$robe&

An RFLP occurs hen the si>e o" a detected
"rag#ent !aries beteen indi!iduals& 6ach
"rag#ent si>e is considered an allele, and can
be used in genetic anal%sis&



)he anal%sis o" CN)R alleles continues, but
is no usuall% $er"or#ed b% $ol%#erase
chain reaction (P<R) #ethods&

For e8a#$le, the standard $rotocols "or
DNA "inger$rinting in!ol!e P<R anal%sis
o" $anels o" #ore than a do>en CN)Rs&

PCR 1POL0./R/AS/ CHAIN
R/ACTION2

DNA "inger$rinting too1 huge
strides "orard in both
discri#inating $oer and the
abilit% to reco!er in"or#ation
"ro# !er% s#all (or degraded)
starting sa#$les&

P<R greatl% a#$li"ies the
a#ounts o" a s$eci"ic region o"
DNA, using oligonucleotide
$ri#ers and a ther#ostable
DNA $ol%#erase&


P<R #ethod is
readil% ada$table "or
anal%>ing CN)R loci&

In +=, the F4I has
standardi>ed a set o"
,@ CN)R assa%s "or
DNA t%$ing, and has
organi>ed database
"or "orensic
identi"ication in
cri#inal cases&

=i#ilar assa%s and
databases ha!e been
set u$ in other
countries&&

P<R
VID/O

0 CHRO.OSO./ ANAL0SIS

Recent inno!ations ha!e included the creation o"
$ri#ers targeting $ol%#or$hic regions on the B5
chro#oso#e (B5=)R), hich allos resolution o" a
#i8ed DNA sa#$le "ro# a #ale and "e#ale andEor
cases in hich a di""erential e8traction is not $ossible&

B5chro#oso#es are $aternall% inherited, so B5=)R
anal%sis can hel$ in the identi"ication o" $aternall%
related #ales&

B5=)R anal%sis as $er"or#ed in the =all% 0e#ings
contro!ers% to deter#ine i" )ho#as *e""erson had
sired a son ith one o" his sla!es& It turns out that he
did&

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