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Detection of synthetic oligonucleotides extracted from body fluids or tissues

  • US 5,693,466 A
  • Filed: 01/03/1995
  • Issued: 12/02/1997
  • Est. Priority Date: 01/08/1993
  • Status: Expired due to Term
First Claim
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1. A method of detecting synthetic oligonucleotides that are present in body fluids or tissues, said oligonucleotides being extractable with isobutanol and ethyl ether, the method comprising the steps of:

  • (a) proteolytically digesting a body fluid or tissue sample;

    (b) extracting the proteolytically digested sample with a partitioning agent;

    (c) extracting the aqueous phase in the extracted sample with isobutanol and ethyl ether,(d) drying the remaining solution containing nucleic acids to a pellet and resuspending the nucleic acids into a solution;

    (e) binding the nucleic acids to a hybridization membrane;

    (f) treating the membrane having bound nucleic acids with a labeled oligonucleotide probe that is complementary to and specifically binds to the synthetic oligonucleotide to be detected;

    (g) washing the membrane to remove any labeled oligonucleotide that is not specifically hybridized to nucleic acids bound to the membrane; and

    (h) detecting the presence of the probe on the membrane.

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