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    Joshua Malonzo
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    Real-time polymerase chain reaction (qPCR) is a technique that measures DNA amplification as it occurs in real-time using fluorescent reporters. There are two common methods - SYBR Green qPCR which uses a fluorescent dye that binds double-stranded DNA, and TaqMan probe qPCR which uses a probe with a reporter-quencher pair that separates upon DNA binding. qPCR allows quantification of the initial amount of DNA or RNA in a sample by measuring the threshold cycle (CT) at which fluorescence exceeds background levels during PCR cycling. It has applications in diagnosing pathogens, measuring gene expression, and analyzing infectivity.

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    Real-time polymerase chain reaction (qPCR) is a technique that measures DNA amplification as it occurs in real-time using fluorescent reporters. There are two common methods - SYBR Green qPCR which uses a fluorescent dye that binds double-stranded DNA, and TaqMan probe qPCR which uses a probe with a reporter-quencher pair that separates upon DNA binding. qPCR allows quantification of the initial amount of DNA or RNA in a sample by measuring the threshold cycle (CT) at which fluorescence exceeds background levels during PCR cycling. It has applications in diagnosing pathogens, measuring gene expression, and analyzing infectivity.

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    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
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    5 views20 pages

    RT PCR

    Uploaded by

    Joshua Malonzo
    Real-time polymerase chain reaction (qPCR) is a technique that measures DNA amplification as it occurs in real-time using fluorescent reporters. There are two common methods - SYBR Green qPCR which uses a fluorescent dye that binds double-stranded DNA, and TaqMan probe qPCR which uses a probe with a reporter-quencher pair that separates upon DNA binding. qPCR allows quantification of the initial amount of DNA or RNA in a sample by measuring the threshold cycle (CT) at which fluorescence exceeds background levels during PCR cycling. It has applications in diagnosing pathogens, measuring gene expression, and analyzing infectivity.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
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    REAL-TIME POLYMERASE

    CHAIN REACTION
    (Quantitative PCR)
    MAIN PRINCIPLE
    Polymerase Chain Reaction Real-Time Polymerase Chain Reaction
    (PCR) (qPCR)

    Determines the Presence or Absence of a The amplification product is measured using


    specific DNA sequence measured at the very fluorescent reporters every time it accumulates
    end of amplification
    The reaction is monitored with each cycle of
    amplification
    CT value refers to when enough products exceed
    the threshold over background
    TWO METHODS OF PERFORMING
    REAL TIME-PCR
    PROCEDURE OF SYBR
    GREEN qPCR
    PROCEDURE OF SYBR GREEN qPCR
    PROCEDURE OF SYBR GREEN qPCR
    PROCEDURE OF SYBR GREEN qPCR
    PROCEDURE OF SYBR GREEN qPCR
    PROCEDURE OF SYBR GREEN qPCR
    PROCEDURE OF
    TAQMAN PROBE
    qPCR
    PROCEDURE OF TAQMAN PROBE
    qPCR

    TaqMan Probe
    Consist of two fluorescent protein
    - Reporter Dye
    - Quencher Dye
    PROCEDURE OF TAQMAN PROBE
    qPCR

    The Reporter will emit its signal once


    the two is separated
    PROCEDURE OF TAQMAN PROBE
    qPCR
    PROCEDURE OF TAQMAN PROBE
    qPCR
    PROCEDURE OF TAQMAN PROBE
    qPCR
    MEASUREMENT
    PROCEDURE OF TAQMAN PROBE
    qPCR

    Higher DNA Concentration


    or High Viral Load
    = Low CT Value

    Lower DNA Concentration


    or Low Viral Load
    = High CT Value
    APPLICATIONS OF
    REAL-TIME PCR
    PROCEDURE OF TAQMAN PROBE
    qPCR

    DIAGNOSING PATHOGENS GENE EXPRESSION ANALYSIS


    INFECTIVITY

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