REAL TIME PCR

REAL TIME PCR - Only in GOD We believe 1 REAL TIME PCR& IT’S REAL TIME PCR& IT’S FUNCTIONS IN DIAGNOSIS FUNCTIONS IN DIAGNOSIS By A

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Unformatted text preview: Only in GOD We believe 11/23/11 1 REAL TIME PCR & IT’S REAL TIME PCR & IT’S FUNCTIONS IN DIAGNOSIS FUNCTIONS IN DIAGNOSIS By: A . Qorani 11/23/11 2 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 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 I n t r o d u c t i o n A d v a n t a g e d i s a d v a n t a g e s R e a l T i m e P C R P r o d u c t D e t e c t i o n i n m e d i c i n e C o n c l u s i o n R e f e r e n c e REAL TIME PCR & IT’S FUNCTIONS IN DIAGNOSIS 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 A d v a n t a g e d i s a d v a n t a g e s R e a l T i m e P C R P r o d u c t D e t e c t i o n i n m e d i c i n e C o n c l u s i o n R e f e r e n c e REAL TIME PCR & IT’S FUNCTIONS IN DIAGNOSIS I n t r o d u c t i o n 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… A d v a n t a g e d i s a d v a n t a g e s R e a l T i m e P C R P r o d u c t D e t e c t i o n i n m e d i c i n e C o n c l u s i o n R e f e r e n c e REAL TIME PCR & IT’S FUNCTIONS IN DIAGNOSIS I n t r o d u c t i o n Exponential Linear Plateau Things which are needed for PCR (1) Template DNA that is going to be amplified.this DNA must be pure & all other contaminations like RNAs must be removed by RNase....
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This note was uploaded on 11/23/2011 for the course BIO 100 taught by Professor Robinson during the Fall '08 term at BYU.

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REAL TIME PCR - Only in GOD We believe 1 REAL TIME PCR& IT’S REAL TIME PCR& IT’S FUNCTIONS IN DIAGNOSIS FUNCTIONS IN DIAGNOSIS By A

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