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    CityU HK Protocol PCR Purification

    Uploaded by

    Taufik Rizkiandi
    The document describes two protocols for purifying PCR products using commercial kits. The FavorPrep GEL/PCR Purification Kit protocol involves mixing the PCR product with binding buffer, passing it through a column, washing with ethanol, and eluting the purified DNA. The Wizard SV PCR Clean-Up System similarly binds DNA to a column, washes with ethanol, and elutes purified DNA.

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    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd

    CityU HK Protocol PCR Purification

    Uploaded by

    Taufik Rizkiandi
    5 views2 pages
    The document describes two protocols for purifying PCR products using commercial kits. The FavorPrep GEL/PCR Purification Kit protocol involves mixing the PCR product with binding buffer, passing it through a column, washing with ethanol, and eluting the purified DNA. The Wizard SV PCR Clean-Up System similarly binds DNA to a column, washes with ethanol, and elutes purified DNA.

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    The document describes two protocols for purifying PCR products using commercial kits. The FavorPrep GEL/PCR Purification Kit protocol involves mixing the PCR product with binding buffer, passing it through a column, washing with ethanol, and eluting the purified DNA. The Wizard SV PCR Clean-Up System similarly binds DNA to a column, washes with ethanol, and elutes purified DNA.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    5 views2 pages

    CityU HK Protocol PCR Purification

    Uploaded by

    Taufik Rizkiandi
    The document describes two protocols for purifying PCR products using commercial kits. The FavorPrep GEL/PCR Purification Kit protocol involves mixing the PCR product with binding buffer, passing it through a column, washing with ethanol, and eluting the purified DNA. The Wizard SV PCR Clean-Up System similarly binds DNA to a column, washes with ethanol, and elutes purified DNA.

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    © All Rights Reserved
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    Brand: FAVORGEN BIOTECH CORP.

    Kit: FavorPrep GEL/PCR Purification Kit


    PCR Clean-Up Protocol:
    1.

    Mix 100 L of PCR product mixture with 5 volumes of FADF Buffer in a


    microcentrifuge tube and vortex vigorously.

    2.

    Place a FADF column (provided in commercial kit) into a Collection Tube.

    3.

    Transfer the PCR mixture to the FADF Column, centrifuge for 30 seconds and
    discard the flow-through eluate.

    4.

    Add 750 L of Wash Buffer (ethanol added) to the FADF Column and centrifuge
    for 30 seconds. Discard the flow-through eluate.

    5.

    Recentrifuge for 3 minutes to dry the column.

    6.

    Place FADF Column to a new microcentrifuge tube.

    7.

    Add 40 L of Elution Buffer or ddH2O to the membrane center of the FADF


    Column and stand FADF Column at room temperature for 2 minutes.

    8.

    Centrifuge for 2 minutes to elute the DNA.

    9.

    Store purified DNA at 4oC or -20oC.

    Wizard SV PCR Clean-Up System protocol


    1.

    Add an equal volume of Membrane Binding Solution to PCR mixture.

    2.

    Insert SV minicolumn (provided in commercial kit) into Collection Tube.

    3.

    Transfer dissolved gel mixture or prepared PCR product to a minicolumn


    assembly.

    4.

    Incubate at room temperature for 1 minute.

    5.

    Centrifuge at 16,000 g for 1 minute. Discard flow-through and reinsert


    minicolumn into Collection Tube.

    6.

    Add 700 L of Membrane Wash Solution (ethanol added). Centrifuge at 16,000


    g for 1 minute. Discard flow through and reinsert minicolumn into a Collection
    Tube.

    7.

    Repeat Step 6 with 500 L of Membrane Wash Solution. Centrifuge at 16,000


    g for 5 minutes.

    8.

    Empty the Collection Tube and re-centrifuge the column assembly for 1 minute
    with the microcentrifuge lid open (or off) to allow evaporation of any residual
    ethanol.

    9.

    Carefully transfer minicolumn to a clean 1.5 mL microcentrifuge tube.

    10. Add 50 L of nuclease-free water to the minicolumn.


    11. Incubate at room temperature for 1 minute. Centrifuge at 16,000 g for 1
    minute.
    12. Store purified DNA at 4 oC or 20 oC.

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