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    DNA extraction (Zymo kit)

    Tsu-Chun Hung

    Published: 2022-06-14 DOI: 10.17504/protocols.io.kxygxz11ov8j/v1
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    Abstract

    DNA extraction (Zymo kit)

    Steps

    Sample Collection

    1.

    Add 200µL of 1mm beads to 1.5ml enppendorf tube

    DNA purification

    1.
    1.
    2.
    2.

    Add 200µL of 0.5mm beads to 1.5ml enppendorf tube

    3.

    Add 600µL of lysis solution to 1.5ml enppendorf tube

    4.

    Collect 9-10µg of sample to 1.5ml enppendorf tube

    Sample crush

    5.

    Put 1.5ml eppendorf tube in the mixmill for sample crush; at condition: 30 rpm/s, for 4mins 0h 4m 0s

    Centrifugation

    6.

    Put 1.5ml eppendorf tube in centrifuge for centrifugation, at this condition:17.0x g,25°C

    7.

    Add 600µL of DNA binding buffer to 1st well of 96 deep well plate

    7.1.

    Add 25µL of magnetic beads to 1st well of 96 deep well plate

    Note
    Shake the bottle and pipetting before using magnetic beads

    7.2.

    Add 200µL of sample (lysate) from 15ml centrifuge microtube to 1st well of 96 deep well plate

    Note
    Usally add last

    8.

    Add 500µL of Wash 1 to 2nd well of 96 deep well plate

    9.

    Add 500µL of Wash 2 to 3rd well of 96 deep well plate

    10.

    Add 500µL of Wash 2 to 4th well of 96 deep well plate

    11.

    Add 100µL of water to 5th well of 96 deep well plate

    12.

    Put in the automated DNA extraction machine

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