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Lambda DNA Extraction Protocol

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  • Extract sample with 500 l Phenol-Chloroform-Isoamyl Alcohol (PCI, 25:24:1) and centrifuge for 5 minutes at 1500 x g to separate the phases.
  • Repeat step 8 in the same PLG 15 ml tube.
  • Using the same PLG 15 ml tube again, extract once with 500 l Chloroform-Isoamyl Alcohol (CI, 24:1) and centrifuge for 5 minutes at 1500 x g to separate the phases.
  • Transfer final aqueous phase to a 1.5 ml microcentrifuge tube, add 45 l 3 M Sodium Acetate, pH 5.2 and 500 l 100% Isopropanol, mix and incubate at room temperature for 15 minutes.
  • Centrifuge 20 minutes at 12,000 x g or more, discard the supernatant and wash pellet two times with 70% Ethanol prior to drying.
  • Resuspend the DNA in 2050 l TE (10 mM Tris-Cl, 1 mM EDTA, pH 8.0).
  • References
    1. Current Protocols in Molecular Biology, Volume I. (1989) Ausubel, F.M. et al. (eds.). John Wiley and Sons, New York, NY pp.1.12.1 - 1.13.10.

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