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Linear amplification of specific nucleic acid sequences

  • US 6,743,605 B1
  • Filed: 06/24/1998
  • Issued: 06/01/2004
  • Est. Priority Date: 06/24/1998
  • Status: Expired due to Term
First Claim
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1. A process for linearly amplifying a specific nucleic acid sequence comprising the steps of:

  • 1) providing;

    said specific nucleic acid sequence, initial primers or nucleic acid constructs comprising two segments, (A) a first segment (i) being sufficiently complementary to a first portion of said specific nucleic acid sequence to bind or hybridize thereto and (ii) which provides for template-dependent extension, and (B) a second segment being (i) substantially non-identical to said first segment, (ii) substantially identical to a second portion of said specific nucleic acid sequence, (iii) which binds to a complementary sequence of said second segment under isostatic or limited cycling conditions after a first complementary copy of said specific nucleic acid sequence has been produced by template dependent extension of said first segment, and (iv) thereby providing for subsequent binding of a first segment of another said initial primer or nucleic acid construct to said first portion of said specific nucleic acid sequence under isostatic or limited cycling conditons, such that after primer extension, a second complementary copy is produced and displaces said first complementary copy; and

    substrates, buffer and a template-dependent polymerizing enzyme; and

    2) incubating said specific nucleic acid sequence and said initial primers or nucleic acid constructs in the presence of said substrates, buffer and template-dependent polymerizing enzyme under isostatic or limited cycling conditions;

    thereby linearly amplifying said specific nucleic acid sequence.

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