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Probes labeled with energy transfer coupled dyes

  • US 6,544,744 B1
  • Filed: 01/20/2000
  • Issued: 04/08/2003
  • Est. Priority Date: 02/01/1994
  • Status: Expired due to Fees
First Claim
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1. A method of identifying and distinguishing different deoxyribonucleic acids produced in chain termination sequencing reactions, said method comprising:

  • (a) producing a first, a second, a third, and a fourth set of different size deoxyribonucleic acids complementary to a deoxyribonucleic acid template using a nucleic acid polymerase, wherein;

    said first set consists essentially of different size deoxyribonucleic acids covalently bonded to a first fluorescent energy-transfer label and terminates with a chain terminator comprising a nitrogenous base complementary to adenine, said second set consists essentially of different size deoxyribonucleic acids covalently bonded to a second fluorescent energy-transfer label and terminates with a chain terminator comprising a nitrogenous base complementary to guanine, said third set consists essentially of different size deoxyribonucleic acids covalently bonded to a third fluorescent energy-transfer label and terminates with a chain terminator comprising a nitrogenous base complementary to thymine, and said fourth set consists essentially of different size deoxyribonucleic acids covalently bonded to a fourth fluorescent energy-transfer label and terminates with a chain terminator comprising a nitrogenous base complementary to cytosine, provided that said first fluorescent energy-transfer label consists of a first specific chemical structure comprising a first fluorescent donor and a first fluorescent acceptor covalently bonded to a first backbone chain, with energy transfer from said first donor to said first acceptor, said second fluorescent energy-transfer label consists of a second specific chemical structure comprising a second fluorescent donor and a second fluorescent acceptor covalently bonded to a second backbone chain, with energy transfer from said second donor to said second acceptor, said third fluorescent energy-transfer label consists of a third specific chemical structure comprising a third fluorescent donor and a third fluorescent acceptor covalently bonded to a third backbone chain, with energy transfer from said third donor to said third acceptor, and said fourth fluorescent energy-transfer label consists of a fourth specific chemical structure comprising a fourth fluorescent donor and a fourth fluorescent acceptor covalently bonded to a fourth backbone chain, with energy transfer from said fourth donor to said fourth acceptor, further provided that the absorption wavelengths of said first donor, said second donor, said third donor, and said fourth donor are substantially the same and said first acceptor, said second acceptor, said third acceptor, and said fourth acceptor, each have different emission wavelengths;

    (b) separating said different size deoxyribonucleic acids produced in said step (a); and

    (c) identifying and distinguishing said different size deoxyribonucleic acids complementary to said deoxyribonucleic acid template separated in said step (b) by irradiating with light absorbed by said first donor, said second donor, said third donor, and said fourth donor; and

    detecting light emission from said first acceptor, said second acceptor, said third acceptor, and said fourth acceptor.

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