Prepared by wasim sir 9377031668

DNA : Structure of Polyucleotide !"ai

DNA Polymer of deoxyribonucleotides
Nucleoside = Nitrogenous base + Pentose sugar (linked through N
glycosidic bond)
Examle adenosine! deoxyadenosine! cytidine! etc"
Nucleotide = Nucleoside + Phoshate grou (linked
through phosphodiester bond)
#any nucleotides link together through $% &% phosphodiester bond to
form olynucleotide chain (as in DNA and 'NA)"
(n course of formation of olynucleotide chain! a hoshate moiety remains
free at &% end of ribose sugar (&% end of olymer chain) and one )*+ grou
remains free at $% end of ribose ($% end of olymer chain)"
Double Helix Model for the Structure of DNA
,cientists in-ol-ed
o Friedrich Meischer .irst identified DNA as an acidic substance
resent in nucleus and named it as /Nuclein0
o Wilkins and Franklin Produced 1)ray diffraction data for DNA
structure
o Watson and Crick Proosed double helix structure model for
DNA based on 1)ray diffraction data
o Erin Chargaff Proosed that in ds DNA! ratios A23 and 425
remain same and are e6ual to one
Features of double helix structure of DNA!
(n a DNA! t7o olynucleotide chains are coiled to form a helix" ,ugar)
hoshate forms backbone of this helix 7hile bases ro8ect in 7ards to each
other"
4omlementary bases air 7ith each other through hydrogen bond" Purines
al7ays air 7ith their corresonding yrimidines" Adenine airs 7ith
thymine through t7o hydrogen bonds 7hile guanine airs 7ith cytosine
through three hydrogen bonds"
o 3he helix is right)handed"
Pitch $"9 nm
:; b in each turn
o 3he lane of one base air stacks o-er the other in a double helix"
3his ro-ides stability to the helix along 7ith hydrogen bonding"
Pac#a$i$ of DNA %eli&
"ackaging of DNA Helix
Distance bet7een t7o consecuti-e base airs in a DNA = ;"$9 nm =
;"$9 < :;
=
m
3otal number of base airs in a human DNA = >"> < :;
=
b
3otal length of human DNA = ;"$9 < :;
=
< >"> < :;
=
= ? @"@ m
@"@ m is too large to be accommodated in the nucleus (:;
>
m)"
*rganisation of DNA in rokaryotes2
o 3hey do not ha-e nucleus" DNA is scattered"
o (n certain regions called nucleoids! DNA (negati-ely charged) is
organised in large loos and is held by some roteins (ositi-ely
charged)"
*rganisation of DNA in eukaryotes2
o 3hey ha-e ositi-ely charged basic roteins called histones (ositi-e
and basic due to resence of ositi-e and basic amino acid residues!
lysine and arginine)"
o +istone octamer Anit of eight molecules of histone
o DNA (negati-ely charged) 7inds around histone octamer (ositi-ely
charged) to form nucleosome"
o : nucleosome has arox" @;; b of DNA"
o Nucleosomes in a chromatin resemble beads resent on strings"
o Beads on string structure in chromatin are further ackaged to form
chromatin fibres! 7hich further coil and condense to form
chromosomes during metahase"
o Non)histone chromosomal roteins Additional set of roteins
re6uired for ackaging of chromatin at higher le-el
'rasformi$ priciple( %ers"ey ad !"ase
e&perimets( ) Properties of $eetic material

Disco#ery of DNA as a $enetic Material
3hough rinciles of inheritance and disco-ery of chromosomes in nucleus
7ere achie-ed long time back! there 7as confusion about 7hich molecule
acted as genetic material"
%ransfor&ing "rinciple
5riffith erformed exeriments 7ith the bacteria Streptococcus
pneumoniae" 3his bacterium has t7o strains , strain and ' strain"
S strain 'acteria ( strain 'acteria
o Produce smooth colonies on
culture late
o Produce rough colonies on
culture late
o +a-e a olysaccharide coat
o Do not ha-e a olysaccharide
coat
o Cirulent (causes neumonia)
o Non)-irulent (does not cause
neumonia)
5riffith0s exeriment
Di-e ' strain in the resence of heat)killed , strain roduce -irulence
because some ho7 ' strain bacteria is transformed by heat)killed , strain
bacteria" +ence! it 7as concluded that there must be transfer of genetic
material"
'ioche&ical Nature of %ransfor&ing Material
A-ery! #cDeod! and #c4arthy 7orked to determine the biochemical
nature of genetic material resonsible for transformation"

3his suggests that DNA has to be the genetic material"

Hershey and Chase Experi&ent to Confir& DNA as the $enetic Material
+ershey and 4hase 7orked on bacteriohages (-iruses that infect bacteria)"
Ehen a bacteriohage infects a bacterium! the -iral genetic material gets
attached 7ith the bacterial genetic material and bacteria then treats the -iral
genetic material as its o7n to synthesise more -iral articles"
+ershey and 4hase 7orked to disco-er 7hether it 7as a rotein or DNA
that entered the bacteria from -irus"
3hey labelled some hages 7ith radioacti-e sulhur and the others 7ith
radioacti-e hoshorus"
3hese radioacti-e hages 7ere used to infect E. coli"
E.coli 7as then blended and centrifuged to remo-e -iral articles"
(t 7as obser-ed that bacteria 7ith radioacti-e DNA 7ere radioacti-e 7hile
those 7ith radioacti-e roteins lost their radioacti-ity"
3his sho7ed that it is the DNA that enters the bacteria from -iruses and not
roteins" +ence! it 7as concluded that DNA is the genetic material"
"roperties of the $enetic Material
(t should be able to relicate (dulicate to roduce its identical coy)"
(t should be chemically and structurally stable"
(t should ha-e scoe for changes that are essential for e-olution"
(t should follo7 the #endelian rinciles of inheritance"
Difference bet7een DNA and 'NA2
DNA (NA
o +as deoxyribose sugar o +as ribose sugar
o &)methyl uracil (thymine) is
resent"
o Aracil is resent in lace of
thymine"
o #ostly DNA acts as
the genetic material"
o 'NA acts as a messenger and
adator" (t acts as a genetic
material in some -iruses"
o DNA is stable"
o Presence of @% *+ grou at
e-ery nucleotide makes 'NA
labile and easily
biodegradable"
o 4hemically less reacti-e!
mutates slo7ly
o #utation in 'NA is faster"
o DNA re6uires 'NA for
rotein synthesis"
DNA F 'NA FProtein
o 'NA directly codes for
roteins"
Why DNA is &ore stable than (NA)
(n 'NA! a @% *+ grou is resent at e-ery nucleotide" 3his makes 'NA
unstable and degradable"
Presence of thymine in lace of uracil confers additional stability to DNA"
'NA being a biocatalyst is more reacti-e"
DNA is double)stranded ha-ing comlementary strand! 7hich resists the
changes by reair mechanism"
DNA *eplicatio wit" +&perimetal Proof ,ac"iery
ad +-ymes ./ol/ed
What is DNA (eplication)
DNA relication is the henomenon in 7hich a dulicate coy of DNA is
synthesised"
(n relication! t7o strands of the DNA helix searate and each strand acts
as a temlate for synthesising ne7 comlementary strands"
After comletion of relication! the t7o coies so roduced 7ill ha-e one
arental and one ne7ly synthesised strand" 3his scheme of relication is
called semi)conser-ati-e relication"
Experi&ent to "ro#e %hat DNA (eplicates Se&i*Conser#ati#ely
Performed by #esselson and ,tahl
E.coli 7as gro7n in a medium containing hea-y isotoe
:&
N as the nitrogen
source"

:&
N 7as incororated into ne7ly synthesised DNA as 7ell and the DNA
became hea-y DNA"
+ea-y DNA molecule can be differentiated from normal DNA by density
gradient centrifugation using cesium chloride as the gradient"
3hen! cells 7ere again transferred into a medium 7ith
:9
N as nitrogen
source" ,amles 7ere taken from this media and their DNA 7as extracted"
E .coli di-ides e-ery @; minutes" 3herefore! the DNA extracted after @;
minutes had a hybrid density"
DNA extracted after 9; minutes had e6ual amount of hybrid and light
intensities"
3his imlies that the ne7ly synthesised DNA obtained one of its strands
from the arent" 3hus! relication is semi)conser-ati-e"
Mechanis& of DNA (eplication
'elication occurs in , hase of cell cycle"
EnGyme in-ol-ed ) DNA olymerase (DNA deendent DNA olymerase)
'elication re6uires energy"
,ource of energy Deoxyribonucleoside trihoshates (DN3Ps)
DN3Ps ha-e dual urose Act as substrates and ro-ide energy also
'elication initiates at secific regions in DNA called origin of relication"
DNA olymerase olymerises a large number of nucleotides in a -ery short
time"
During the course of relication! t7o arent strands do not comletely
oen! but a small oening forms in 7hich relication occurs" 3his small
oening forms a relication fork"
DNA olymerase can olymerise only in one direction that is '"
3herefore! relication occurs smoothly at to end of DNA" (continuous
relication! but occurs discontinuously at to end)
3he discontinuous fragments so formed are 8oined by DNA ligase"
'rascriptio 0it 111 Structure ad its *elatios"ip
wit" a 2ee
%ranscription
3ranscrition is the rocess of formation of 'NA molecules from the DNA"
During transcrition! only a segment of DNA from only one of the strands
articiates"
Both strands are not coied during transcrition because2
o (f both strands get transcribed at the same time since the se6uences
of amino acid 7ould be different in both (due to comlementarity)!
then t7o 'NA molecules 7ith different se6uences 7ill be formed!
7hich in turn gi-e rise to t7o different roteins" 3herefore! one
DNA 7ould end u gi-ing rise to t7o different roteins"
o 37o 'NA molecules so formed 7ill be comlementary to each
other! hence 7ould end u forming a double)stranded 'NA lea-ing
the entire rocess of transcrition futile"
%ranscriptional +nit
A transcritional unit has rimarily three regions2
o Promoter #arks the beginning of transcritionH 'NA olymerase
binds here
o ,tructural gene Part of the DNA that is actually transcribed
o 3erminator #arks the end of transcrition
%e&plate Strand and Coding Strand
EnGyme in-ol-ed in transcrition! 'NA olymerase (DNA deendent 'NA
olymerase)! catalyses in only one direction i"e"! &% to $%"
3herefore! the strand 7ith olarity $% F &% acts as a temlate (3emlate
,trand)"
3he strand 7ith olarity &% F $% acts as coding strand (7hich is a misnomer
since it does not code for anything)" 4oding strand has se6uence similar to
'NA formed after transcrition excet for the change that thymine is
resent instead of uracil"
$ene
3he DNA se6uence 7hich codes for t'NA or r'NA molecule defines a
gene"
4istron ,egment of DNA that contains the genetic code for a single
olyetide
3he structural genes could be of t7o tyes2
o #onocistronic (mostly in eukaryotes)
o Polycistronic (mostly in rokaryotes)
#onocistronic genes ha-e t7o arts2
o Exon ,e6uences that code for a articular character and is
exressed in a matured and rocessed m'NA
o (ntron (nterruting se6uences that do not aear in a mature and
rocessed m'NA
'egulatory genes ,e6uences that do not code for anything! but ha-e
regulatory functions
'ypes of *NA ) 'rascriptio Process

%ypes of (NA
m'NA (messenger 'NA) (t ser-es as a temlate for rotein synthesis"
DNA is transcribed to form an m'NA! 7hich in turn is translated to form
rotein" I4entral dogma of molecular biologyJ
t'NA (transfer 'NA) (t brings amino acids during translation and reads
the genetic code"
r'NA (ribosomal 'NA) 3hese are the 7ork benches of translation" 3hey
lay a structural and catalytic role during translation"
%ranscription "rocess
3ranscrition has three stes initiation! elongation! and termination"
(nitiation2
o 'NA olymerase binds 7ith the romoter to initiate the rocess of
transcrition"
o Association 7ith initiation factor (K) alters the secificity of 'NA
olymerase to initiate the transcrition"
Elongation!
o 'NA olymerase uses nucleotide trihoshate as substrate! and
olymerisation occurs according to comlementarity"
3ermination2
o 3ermination occurs 7hen termination factor (P) alters the secificity
of 'NA olymerase to terminate the transcrition"
o As the 'NA olymerase roceeds to erform elongation! a short
stretch of 'NA remains bound to the enGyme" As the enGyme
reaches the termination region! this nascent 'NA falls off and
transcrition is terminated"
Co&plexities Associated ith %ranscription
(n rokaryotes2
o 3here is no clear demarcation bet7een cytosol and nucleus"
3herefore! translation can begin e-en before transcrition is
comleted" 3hus! in rokaryotes! transcrition and translation are
couled"
(n eukaryotes2
o 3hree different kinds of 'NA olymerases are resent"
'NA olymerase ( transcribes r'NA"
'NA olymerase (( transcribes hn'NA (m'NA recursor)"
'NA olymerase ((( transcribes t'NA! sn'NA! and sr'NA"
o 3he recursor of m'NA! i"e" hn'NA! contains both introns and
exons" (ntrons are remo-ed and exons are 8oined by a rocess called
slicing"
o 4aing (n this! methyl guanosine trihoshate is added to the
&% end of hn'NA"
o 3ailing (n this! adenylate residues are added to the $% end of
hn'NA"
o Ehen hn'NA is fully rocessed! it is kno7n as m'NA! 7hich is
transorted out of the nucleus to get translated"
2eetic !ode ad Study of ,utatios
$enetic Code
5enetic code directs the se6uence of amino acids during the synthesis of
roteins"
5eorge 5amo7 roosed that if @; amino acids are to be coded by 9 bases!
then the code should be made u of three nucleotides" 9
$
= >9 (9
@
= :>)!
7hich is less than @;H so! the codon 7as roosed to be trilet"
+ar 5obind Lhorana de-eloed a chemical method to synthesise 'NA
molecules 7ith defined combination of bases"
Nirenberg de-eloed cell)free systems for rotein synthesis! 7hich heled
the code to be decihered"
3he enGyme kno7n as ,e-ero *choa enGyme (olynucleotide
hoshorylase) heled to olymerise 'NA 7ith defined se6uences in a
temlate indeendent manner"
(t finally ga-e rise to the checker)board for genetic code"
,alient features of genetic code2
o 4odon is trilet" 9
$
= >9 (>: codons code for amino acids 7hile $ are
sto codons)
o *ne codon codes for a single secific amino acid" 4odons are
unambiguous"
o 4odons are degenerate since some amino acids are coded by more
than one codon"
o 5enetic code is uni-ersal" : codon codes for same amino acid in all
secies"
o 4odons are read continuous" 3hey lack unctuations"
o AA5 has dual functions 4odes for #ethionine and acts as a start
codon
Effects of Mutations on $enetic Code
#utations include insertions! deletions! and rearrangements"
#utation results in changed henotye and diseases such as sickle cell
anaemia" (4hange 5luF Cal in gene coding for beta globin chain of
haemoglobin) ,uch mutations are called ,oint &utations"
(nsertion or deletion of a single base air disturbs the entire reading frame
in m'NA" ,uch mutations are called fra&eshift &utations"
.rameshift mutations hold the roof of the fact that codon is trilet because if 7e
insert three or multile of three bases follo7ed by the deletion of same number of
bases! then the reading frame 7ill remain unaltered"
Structure of t*NA3 Process of 'raslatio3 *e$ulatio of
2ee +&pressio
t(NA
t'NA is an adater molecule" *n one hand! it reads the genetic code and on
the other hand! it binds to secific amino acids"
t'NA has an anticodon loop that has bases comlementary to the m'NA
code and an a&ino acid acceptor end 7here it binds to the corresonding
amino acid"
(nitiation t'NA 3his t'NA is essential for initiation of translation and
has AA5 in anticodon loo and #et in amino acid accetor end"
3here are no t'NAs for sto codons"
%ranslation
3he m'NA contains the genetic information! 7hich is translated into the
amino acid se6uence 7ith hel of t'NA" Amino acids are olymerised to
form a olyetide"
Amino acids are 8oined by etide bond"
.irst of all! charging of t'NA (amino)acylation of t'NA) takes lace" (n
this! amino acids are acti-ated in the resence of A3P and are linked to
their corresonding t'NA"
'ibosomes are the 7orkbenches for translation" 'ibosomes ha-e @
subunits2 a large subunit and a small subunit"
,maller subunit comes in contact 7ith m'NA to initiate the rocess of
translation"
3ranslational unit in an m'NA is the region flanked by start codon and sto
codon"
Antranslated regions (A3') are the regions on m'NA that are not
themsel-es translated! but are re6uired for efficient translation rocess"
3hey may be resent before start codon (&%A3') or after sto codon
($% A3')"
(nitiator t'NA recognises the start codon" ((nitiation)
3hen t)'NA)amino acid comlexes bind to their corresonding codon
on the m'NA and base airing occurs bet7een codon on m'NA and t'NA
anticodon"
t'NA mo-es from codon to codon on the m'NA and amino acids are
added one by one" (Elongation)
'elease factor binds to sto codon to terminate the translation"
(3ermination)
(egulation of $ene Expression
'egulation of gene exression could be exerted at follo7ing le-els"
o 3ranscritional le-el (follo7ing of rimary transcrits)
o Processing le-el (slicing)
o 3ransort of m'NA from nucleus to cytolasm
o 3ranslational le-el
(n addition! metabolic! hysiological! or en-ironmental conditions regulate
the exression of genes"
Exression of genes coding for enGymes is re6uired only 7hen substrate for
that enGyme is a-ailable"
.or examle2
Dactose 5lucose + 5alactose
E.coli synthesises beta)galactosidase! only 7hen lactose is a-ailable"
'egulation in rokaryotes
o 5ene exression is regulated by controlling the rate of
transcritional initiation"
o 3he acti-ity of 'NA olymerase at a gi-en romoter is regulated by
accessory roteins" 3he accessory roteins affect the ability of a
romoter to recognise start sites"
o A regulatory rotein could be acti-ator or reressor"
o Accessibility of romoter is also affected by oerators" *erator is
the region located ad8acent to romoter"
o Each oeron has a secific oerator and a secific reressor"
o Asually oerator binds to a reressor rotein"
*e$ulatio of 4ac 5pero
Lac -peron
*eron An arrangement 7here a olycistronic gene is regulated by a
common romoter and regulatory genes
Lac oeron! trp oeron! his oeron! val oeron are the examles of such
systems"
3he elucidation of lac oeron as a transcritionally acti-e system 7as first
done by geneticist Macob and biochemist #onod"
5enes constituting lac oeron2
$ene Nature Function
i gene (nhibitor (t codes for reressor of lac oeron"
z gene ,tructural (t codes for N)galactosidase"
Dactose 5alactose + 5lucose
ygene ,tructural (t codes for ermease! 7hich increases the ermeability
of cell to N)galactosidase"
agene ,tructural (t codes for transacetylase"
All genes in-ol-ed in lac oeron are re6uired for metabolism of lactose"
.nducer Dactose acts as an inducer for lac oeron since it regulates the
s7itching on and off of the oeron"
(f lactose is ro-ided to the gro7th media of bacteria in absence of any
other carbon source! then it is transorted inside the cells by ermease"
.or ermease to be resent and lactose to enter inside the cells! lo7 le-el of
exression of lac oeron must be resent all the time"
(egulation in Absence of .nducer
(n absence of inducer! i gene transcribes to synthesise reressor m'NA!
7hich translates to form reressor"
3his reressor binds 7ith the oerator region of oeron and re-ents 'NA
olymerase to transcribe genes z! y! and a (negati-e regulation)"
3herefore! in absence of the roducts of these genes! metabolism of lactose
ceases"
(egulation in "resence of .nducer
(nducer binds 7ith the rotein roduct of gene i (reressor) and inacti-ates
it"
3his inacti-ated reressor is unable to bind to oerator region so that 'NA
olymerase transcribing the oeron enGyme and z! y! and a genes synthesise
their resecti-e m'NA! 7hich in turn gets translated to form N)
galactosidase! ermease! and transacetylase"
(n resence of all these enGymes! the metabolism of lactose roceeds in a
normal manner"
%uma 2eome Pro6ect 7%2P8
Moint -enture of A, deartment of energy and National (nstitute of +ealth
(N(+)H later 8oined by Eelcome 3rust (AL)
Daunched in :==;! comleted in @;;$
3his ro8ect 7orked to7ards the determination of comlete DNA se6uence
of humans"
DNA is the storehouse of genetic information and determining its se6uence
of base airs can sol-e many medical! agricultural! en-ironmental! and
e-olutionary mysteries"
(elationship of H$" ith 'ioinfor&atics
+uman genome (genome refers to the totality of genes that are resent in a
human being) contains $ < :;
=
base airs"
4ost of se6uencing : b = A, O $
4ost of se6uencing $ < :;
=
b = A, O = billion
Enormous se6uence data so generated 7ould ha-e re6uired $$;; books
containing :;;; ages each 8ust for a human genome"
+ence! for storing! retrie-ing! and analysing this enormous data! a ne7
branch of biology has been de-eloed kno7n as bioinformatics"
5enomes of many non)human models such as bacteria!
yeast! Caenorhabditis elegans!Drosophila! lants (rice and Arabidopsis)
ha-e also been se6uenced"
Methods to .dentify $enes
37o methods identifying E,3s (Exressed se6uence 3ags) and se6uence
annotation
E,3s As the name suggests! this refers to the art of DNA that
is exressed! i"e" transcribed! as m'NA and translated into roteins
thereafter" (t basically focuses on se6uencing the art denoting a gene"
Annotation (n this aroach! entire genome (coding + non)coding) is
se6uenced and later on function is assigned to each region in the genome"
$eno&e Se/uencing
DNA from the cells is isolated and is randomly broken into fragments of
smaller siGes"
3hese fragments are cloned into suitable host using -ectors"
4loned fragments amlify in the host" Amlification facilitates an easy
se6uencing"
4ommon -ectors used BA4 (Bacterial artificial chromosomes) and PA4
(Peast artificial chromosomes)
4ommon hosts Bacteria and yeasts
Automated se6uencers are used to se6uence these smaller fragments
(,anger se6uencing)"
3he se6uences so obtained are arranged based on o-erlaing regions
7ithin them (alignment)"
Alignment of the se6uences is also done automatically by comuter
rograms"
3hen these se6uences are annotated and assigned to each chromosome"
"reparation of $enetic and physical &aps on $eno&e
@ methods are used restriction olymorhism and microsatellites
'estriction olymorhism ,ecialiGed enGymes called restriction
endonucleases are used to cut the genome at secialiGed sites called
restriction endonuclease recognition site and mas are reared based on it"
#icrosatellites 3hese are reetiti-e DNA se6uences"
-bser#ations fro& H$"
+uman genome contains $ < :;
=
($:>9"Q million) nucleotide bases"
An a-erage gene consists of $;;; bases" +o7e-er! the siGe of genes -aries"
Dargest gene is dystrohin (@"9 m bases)"
3otal number of genes in human genome $;!;;;
*-er &;R of the disco-ered genes ha-e unkno7n functions"
Dess than @R of genome is coding"
Darge ortion of genome consists of reeating se6uences"
'eetiti-e se6uences ha-e no coding function" 3hey are reeated o-er
hundred to thousand times" 3hey may ha-e a role in e-olution! chromosome
structure! and dynamics"
4hromosome 7ith most genes 4hromosome : (@=>S)
4hromosome 7ith fe7est genes 4hromosomes P (@$:)
,NPs (single nucleotide olymorhism) occur at about :"9 million locations
in human DNA" 3hey are belie-ed to ha-e significance in exlaining
diseases and e-olutionary history of human beings"
DNA 9i$erpriti$
.ntroduction
DNA fingerrinting is a method for comaring the DNA se6uences of any
t7o indi-iduals"
=="=R of the base se6uences in all human beings are identical" (t is the
remaining ;":R that makes e-ery indi-idual uni6ue"
(t is a really difficult and time)consuming task to se6uence and comare all
$ < :;
=
bases in t7o indi-iduals" ,o! instead of considering the entire
genome! certain secific regions called reetiti-e DNA se6uences are used
for comarati-e study"
'asis of DNA Fingerprinting
'eetiti-e DNA is searated from bulk genomic DNA since it aears as a
distinct eak during density gradient centrifugation"
#a8or eak2 .ormed by bulk DNA
,maller eak2 ,atellite DNA
,atellites are of t7o tyesTmicro)satellites and mini satellites! deending
uon the base comosition! length of segment and the number of reetiti-e
units"
,atellites do not code for roteins! but ha-e a ma8or role to lay in DNA
fingerrinting"
Polymorhism is actually a result of mutation" A germ cell mutation (7hich
can ass on to the next generation through sexual reroduction) gi-es rise
to olymorhism in oulations"
(n other 7ords! an inheritable mutation if obser-ed in higher fre6uencies in
a oulation is kno7n as olymorhism"
Polymorhisms arise normally in non)coding se6uences because mutations
in non)coding se6uences do not affect an indi-idual0s reroducti-e ability"
Methodology of DNA fingerprinting
CN3' (-ariable number of tandem reeats) are satellite DNAs that sho7
high degree of olymorhism"
CN3's are used as robes in DNA fingerrinting"
.irst of all! DNA from an indi-idual is isolated and cut 7ith restriction
endonucleases"
.ragments are searated according to their siGe and molecular 7eight on
gel electrohoresis"
.ragments searated on electrohoresis gel are blotted (immobilised) on a
synthetic membrane such as nylon or nitrocellulose"
(mmobilised fragments are hybridised 7ith a CN3' robe"
+ybridised DNA fragments can be detected by autoradiograhy"
CN3's -ary in siGe from ;": to @; kb"
+ence! in the autoradiogram! band of different siGes 7ill be obtained"
3hese bands are characteristic for an indi-idual" 3hey are different in each
indi-idual! excet identical t7ins"
Applications of DNA Fingerprinting
DNA fingerrinting is 7idely used in forensics since e-ery DNA of e-ery
tissue from an indi-idual has the same degree of olymorhism"
DNA fingerrinting forms the basis of aternity testing since a child
inherits olymorhism from both its arents"
(t can be used for studying genetic di-ersity in a oulation and e-olution"