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  • Polymerase Chain Reaction: Martinez - Ortega - Villegas

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    James Carl Villegas
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    This document provides an overview of polymerase chain reaction (PCR) including its history, components, and primary steps. PCR is an in vitro technique used to amplify a specific region of DNA. It was developed in 1983 by Kary Mullis who received the Nobel Prize in 1993. Key components of PCR include a DNA template, primers, a thermostable DNA polymerase such as Taq polymerase, nucleotides, magnesium chloride, and buffers. The primary steps of PCR are denaturation of the DNA template, annealing of primers to the single-stranded DNA, and extension of the primers by DNA polymerase to replicate the DNA sequence.

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    polymerase chain reaction principles

    Original Title

    Pcr

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    This document provides an overview of polymerase chain reaction (PCR) including its history, components, and primary steps. PCR is an in vitro technique used to amplify a specific region of DNA. It was developed in 1983 by Kary Mullis who received the Nobel Prize in 1993. Key components of PCR include a DNA template, primers, a thermostable DNA polymerase such as Taq polymerase, nucleotides, magnesium chloride, and buffers. The primary steps of PCR are denaturation of the DNA template, annealing of primers to the single-stranded DNA, and extension of the primers by DNA polymerase to replicate the DNA sequence.

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    © All Rights Reserved
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    12 views10 pages

    Polymerase Chain Reaction: Martinez - Ortega - Villegas

    Uploaded by

    James Carl Villegas
    This document provides an overview of polymerase chain reaction (PCR) including its history, components, and primary steps. PCR is an in vitro technique used to amplify a specific region of DNA. It was developed in 1983 by Kary Mullis who received the Nobel Prize in 1993. Key components of PCR include a DNA template, primers, a thermostable DNA polymerase such as Taq polymerase, nucleotides, magnesium chloride, and buffers. The primary steps of PCR are denaturation of the DNA template, annealing of primers to the single-stranded DNA, and extension of the primers by DNA polymerase to replicate the DNA sequence.

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    of 10

    1

    B I O 1 7 1 - 1 L

    POLYMERASE CHAIN
    REACTION
    MARTINEZ

    ORTEGA -

    VILLEGAS

    ABOUT US
    MARTINEZ, MARJORIE
    ORTEGA, VANESSA
    VILLEGAS, JAMES
    BIO171-1L
    FIRST QUARTER
    AY 2016-2017
    PROF. UREAH THEA SEVILLA

    OUTLINE
    Overview
    What is PCR together with a brief introduction

    Principles
    Processes, steps and discussion of the PCR

    Techniques
    Difference approaches in PCR technology

    Applications
    Existing and future applications of PCR technology

    OVERVIEW
    PCR, polymerase chain reaction, is an in-vitro technique for
    amplification of a region of DNA whose sequence is known
    or which lies between two regions of known sequence

    Before PCR, DNA of interest could only be amplified by overexpression in cells and this with limited yield

    HISTORY
    1966, Thomas Brock discovers Thermus Aquaticus, a
    thermostable bacteria in the hot springs of Yellowstone
    National Park

    1983, Kary Mullis postulated the concept of PCR ( Nobel


    Prize in 1993)
    1985, Saiki publishes the first application of PCR ( betaGlobin)
    1985, Cetus Corp. Scientists isolate Thermostable Taq
    Polymerase (from T.Aquaticus), which revolutionized PCR

    COMPONENTS
    DNA TEMPLATE
    DNA template that contains the DNA region (target) to
    amplify

    PRIMERS
    Good primer design is essential for successful reactions.
    The important design considerations are a key to
    specific amplification with high yield.
    ENZYME (Usually Taq DNA Polymerase)
    DNA polymerase with a temperature optimum at around
    70 C. DNA Polymerase should be able to elongate the DNA
    strand with the help of Primers, as the DNA Polymerase can not
    attach to a DNA strand and elongate on its own. It should also be
    heat resistant, so that it can withstand the denaturation process.

    COMPONENTS
    Deoxynucleoside triphosphates
    Adenine, Thymine, Guanine, Cytosine
    Mg2+

    Mg2+ is a cofactor for Taq enzyme and act as a


    catalyst.
    BUFFERS
    Providing a suitable chemical environment for optimum
    activity and stability of the DNA polymerase

    PCR

    POLYMERASE CHAIN REACTION


    IN-VITRO APPLICATION
    GENE AMPLIFICATION

    PRIMARY STEPS
    THESE ARE THE THREE PRIMARY STEPS TO REPLICATE A DNA SEQUENCE USING POLYMERASE
    CHAIN REACTION

    DENATURATION
    DENATURATION
    OF DNA
    TEMPLATE AT
    95 C

    ANNEALING
    TEMPERATURE
    IS LOWERED
    FOR PRIMERS
    TO ANNEAL

    EXTENSION
    TAQ
    POLMERASE
    CATALYZES
    THE PRIMER
    EXTENSION

    10

    THANK YOU!

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