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Chain-terminating nucleotides for DNA sequencing methods

  • US 5,547,859 A
  • Filed: 08/02/1993
  • Issued: 08/20/1996
  • Est. Priority Date: 08/02/1993
  • Status: Expired due to Fees
First Claim
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1. A method for sequencing DNA, comprising:

  • contacting a polymerase selected from the group consisting of T4 polymerase, T2 polymerase, T6 polymerase and E. coli DNA polymerase II, with a primed DNA strand to be sequenced in the presence of dATP, dGTP, dCTP, dTTP, a first chain-terminating nucleotide, a second chain-terminating nucleotide, a third chain-terminating nucleotide and a fourth chain-terminating nucleotide; and

    allowing said contacting to proceed under reaction conditions to maintain polymerase activity for a period of time sufficient to obtain sequencing information, wherein said first chain-terminating nucleotide is 3'"'"'-amino-2'"'"',3'"'"'dideoxy-ATP, said second chain-terminating nucleotide is 3'"'"'-amino-2'"'"',3'"'"'dideoxy-GTP, said third chain-terminating nucleotide is 3'"'"'-amino-2'"'"',3'"'"'dideoxy-CTP and said fourth chain-terminating nucleotide is 3'"'"'-amino-2'"'"',3'"'"'dideoxy-TTP.

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