US 4,971,903 AGranted

Pyrophosphate-based method and apparatus for sequencing nucleic acids

OwnerEdward Hymaninventor

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Abstract

A method for sequencing nucleic acid polymers is provided in which the polymer to be sequenced acts as a template for the production of a complementary polymer by a polymerase enzyme. The template polymer is introduced into a polymerization environment in which production of the complementary polymer will occur if appropriate nucleotides are provided. The nucleotides are then provided to the polymerization environment one at a time in individual feedstocks. If the nucleotide in a feedstock is complementary to the next base in the template polymer, i.e., the unpaired base closest to the growing end of the complementary polymer, polymerization will occur lengthening the complementary polymer and releasing PPi. By separately recovering each feedstock and analyzing it for the presence of PPi, the sequence of the complementary polymer and thus the template polymer is determined.

Claims13

Dependent claims

A method for determining the nucleic acid sequence in a template nucleic acid polymer, comprising

(a) introducing the template nucleic acid polymer into a polymerization environment in which the nucleic acid polymer will act as a template polymer for the synthesis of a complementary nucleic acid polymer when nucleotides are added;
(b) successively providing to the polymerization environment a series of feedstocks, each feedstock comprising a nucleotide selected from among the nucleotides from which the complementary nucleic acid polymer will be formed, such that if the nucleotide in the feedstock is complementary to the next nucleotide in the template polymer to be sequenced said nucleotide will be incorporated into the complementary polymer and inorganic pyrophosphate will be released;
(c) separately recovering each of the feedstocks from the polymerization environment;
and
(d) measuring the amount of inorganic pyrophosphate in each of the recovered feedstocks to determine the identity of each nucleotide in the complementary polymer and thus the sequence of the template polymer.

2 more elements

Dependencies are inferred from the claim text.