Real time PCR and it's functions in diagnosis

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Only

in

GOD

We believe

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REAL TIME PCR & IT’S FUNCTIONS IN DIAGNOSIS

By: A . Qorani

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1:Introduction to PCR 2:Advantages & disadvantages 3:Real Time PCR 4:Detection of products 5:Function in medicine 6:Conclusion 7:Reference

Outline

9/4/2015

Introduction

Advantage disadvantages

Real Time PCR

Product Detection

in medicine

Conclusion

Reference

REAL TIME PCR & IT’S FUNCTIONS IN DIAGNOSIS

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Introduction to PCR

PCR was invented in 1984 by ( Kary mullis ) & he received the Nobel Prize in chemistry in 1993, for his invention.(1)

It revolutionized biological methods specially in molecular cloning in a way that it has

became an inseparable & irreplaceable part of molecular investigations.

Introduction

Advantage disadvantages

Real Time PCR

Product Detection

in medicine

Conclusion

Reference

REAL TIME PCR & IT’S FUNCTIONS IN DIAGNOSIS

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more…

There is three basic steps which are in common with all type of PCR

Thermal denaturation :

In this step DNAs are denatured mostly by temprature about 94˚c & single stranded DNAs are made.

( in some cases It’s done by helicase )

Primer annealing :

In this step Primers are attached to ssDNA by their complementary regions.

Extension or polymerization :

This is done by a temprature resistance polymerase named Taq polymerase which is extracted from Thermus aquaticus.

Advantage disadvantages

Real Time PCR

Product Detection

in medicine

Conclusion

Reference

REAL TIME PCR & IT’S FUNCTIONS IN DIAGNOSIS

Introduction

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PCR phases

Exponential

If 100% efficiency – exact doubling of products. Specific and precise

Linear

High variability. Reaction components are being consumed and PCR products are starting to degrade.

Plateau

End-point analysis. The reaction has stopped and if left for long – degradation of PCR products.

more…

Advantage disadvantages

Real Time PCR

Product Detection

in medicine

Conclusion

Reference

REAL TIME PCR & IT’S FUNCTIONS IN DIAGNOSIS

Introduction

Exponential

Linear

Plateau

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Things which are needed for PCR(1)