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  • 發布時間:2019-04-22 23:35 原文鏈接: TCA沉淀法定量DNA

    1. Dilute radioactive sample to a 100 ml volume

    2. Spot 5 ml of the sample onto the center of a 2.4 cm Whatman GF/C glass-fiber disc.

    3. Mix 5 ml of the sample with 100 ml Salmon sperm DNA (50 mg in 20 mM EDTA).

    4. Add 5 mL ice cold 10% Trichloroacetic acid (TCA). Mix well and incubate on ice for 15 min.

    5. Filter the solution through a separate GF/C glass-fiber disc. 

    6. Wash the filter 6 times with 5 mL ice-cold 10% TCA.

    7. Wash filter once with 5 mL 95% Ethanol.

    8. Dry both filter under a heat lamp.

    9. When dry, count each in a scintillation counter.

    10. The first filter measures total radioactivity. The second filter measures radioactivity incorporated into DNA fragments greater than 20 nucleotides in length.

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