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    Home > Biochemistry News > Biotechnology News > Geogosin-based randomly labeled DNA probes.

    Geogosin-based randomly labeled DNA probes.

    • Last Update: 2020-10-30
    • Source: Internet
    • Author: User
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    1. Mark
    DNA
    Probe Each standard reaction can mark 10ng to 3linear DNA, or a larger amount of DNA, but all ingredients and volumes increase accordingly.(1) DNA probe hot denaturation, boiling 10min, quickly cooled in ice/ethanol more than 5min, to be used.
    (2) take Eppendorf tube (1.5 ml) and place it on ice, add the following and
    reagents
    :
    freshly denatured DNA 1-3
    he6-poly
    nucleotides
    mixture 2 (tube 5)
    d NTP marker with mixed substrate 2 (tube 6)
    plus sterile heavy steamed water to 19
    DNA polymerase I. large fragment (Klenow) 1 (tube 7)
    (3) insulation at 37 degrees C at least 60min, up to 20h.
    (4) boil 5min to terminate the reaction.
    (5) 15000rpm centrifuge 30s and place on ice for use.
    2. Pre-hybridization According to 100cm2 membrane with 20 to 40 ml pre-hybrid solution, pre-hybridization makes the solution in a flowing state.
    prebreeding liquid composition: 5㗓SC
    0.5% (W/V) sealing reagent (tube 11)
    0.1% (W/V) N-12 Sodium alkyl creatinine
    0.02% (W/V)
    SDS
    membrane into a plastic bag, filled with pre-hybrid liquid, removed the gas after sealing, pre-hybridized at 65 degrees C overnight.
    .
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