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Fingerprinting using single specific primers in low stringency polymerase chain reaction conditions

  • US 5,599,674 A
  • Filed: 03/20/1995
  • Issued: 02/04/1997
  • Est. Priority Date: 07/29/1993
  • Status: Expired due to Fees
First Claim
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1. A method to obtain a signature characteristic of the nucleotide sequence of a purified cloned or amplified defined DNA segment which method consists essentially of:

  • subjecting said segment to multiply cycles of replication in the presence of a single primer fully complementary to a portion of at least about ten nucleotides of said segment, wherein said portion is not a repetitive sequence or part of a gene cluster within said DNA segment under conditions of low stringency at 25°

    C.-40°

    C. a primer concentration in the range of 2-10 μ

    M, and a polymerase concentration in the range of 60-200 units/ml to obtain a mixture of replicated DNA molecules which result from said multiplicity of low stringency cycles of replication; and

    determining the size of the replicated DNA molecules in the mixture thus obtaining a signature characteristic of said nucleotide sequence.

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