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REAL TIME PCR

REAL TIME PCR - GOD Only in We believe 1 REAL TIME PCR ITS...

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Only in GOD We believe 11/23/11 1
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REAL TIME PCR & IT’S REAL TIME PCR & IT’S FUNCTIONS IN DIAGNOSIS FUNCTIONS IN DIAGNOSIS By: A . Qorani 11/23/11 2
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1:Introduction to PCR 1:Introduction to PCR 2:Advantages & disadvantages 2:Advantages & disadvantages 3:Real Time PCR 3:Real Time PCR 4:Detection of products 4:Detection of products 5:Function in medicine 5:Function in medicine 6:Conclusion 6:Conclusion 7:Reference 7:Reference Outline 11/23/11 3 11/23/11 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.  11/23/11 4 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.  11/23/11 5 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 in linear view Cycle # [DNA] PCR phases 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. 11/23/11 6 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) Template  DNA  that  is  going  to  be  amplified.this  DNA must be pure & all other contaminations like  Cations  that are essential for polymerase activity.  11/23/11 7 more… Advantage disadvantages Real Time PCR Product Detection in medicine Conclusion Reference REAL TIME PCR & IT’S FUNCTIONS IN DIAGNOSIS Introduction Contaminations like RNAs must be removed by  RNase.
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Advantages & disadvantages * The most accurate & feasible technique to       determine the amount & concentration of products.
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