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Polymerase Chain Reaction
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Polymerase Chain Reaction is a biochemical
technique for in vitroDNA amplification.
PCR was first described by Kary Mullis in 1983..
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. a process by which DNA is artificially
rep ca e y repea e cyc es o an n v roDNA polymerase reactions.
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.
There are three major steps in a PCR, which.
. .
.
3. extension 72 C .
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Target DNA Sequence
53 Denaturation
35
Annealing
53
35
53 Extension
5 353
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The thermal cycler (PCR machine).
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The gel electrophoresis appartus
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DNA electrophorogram
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PCR components.
DNA template.
ne or morepr mers.
DNA polymerase (e.g. Taq polymerase).
Deoxynucleotide triphosphates.
.
Divalent cations(Mg+2
).
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UsedtoidentifyapersonbycomparinghisorherDNA
withthe
DNA
in
a
iven
sam le.
patternof
DNA
fragments
generated
by
restriction
' '
.
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Electrophoresis of PCR-amplified DNA fragments.
Genetic relationships can be determined bycomparing DNA fingerprints.
This technique can also be used to determine
evolutionar relationshi s between or anisms.
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Targetgenescanbeamplifiedthensequencedto
detectmutations
(hereditary).
.
sequencecanbeanalyzedbyPCR.
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Cloning genes
,
genefromoneorganismandtheninsertingitinto
.
PCRisusedtoamplifyDNAfragments(genes).
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Archeological analysis of fossil.
PCR,to
analyze
fossil
DNA.
oldDNA
(mammoth).
The
analysis
of
Egyptian
mummies
.
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polymorphismexists.
Singlenucleotidepolymorphism.
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uan a vevers onso areuse omon orgeneexpression.
mRNAisisolatedandreversetranscribedintocDNA.
Reversetranscriptase
PCR
(RT
PCR).
RealtimePCR (RTPCR)
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amplify,isolateoridentifyaknownsequencefrom.
reversetranscriptase.
Iswidelyusedinexpressionmapping,determining.
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RealtimePCRisthetechniqueofcollectingdata
throughoutthe
PCR
process
as
it
occurs,
thus
combiningamplificationanddetectionintoasingle
step.
Thismethodusesfluorescentd esand robesto
measuretheamountofamplifiedproductinreal
time.
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