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US 6,255,083 B1Granted
System and methods for nucleic acid sequencing of single molecules by polymerase synthesis
OwnerJohn G. K. Williamsinventor
Life of this patent
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Abstract
This invention relates to improved methods for sequencing and genotyping nucleic acid in a single molecule configuration. The method involves single molecule detection of fluorescent labeled PPi moieties released from NTPs as a polymerase extension product is created.
Claims21
A method of genotyping or sequencing a target nucleic acid comprising the steps of;
- i. immobilizing onto a solid support a complex comprising a target nucleic acid, a primer nucleic acid which complements a region of the target nucleic acid, and at least one nucleic acid polymerase;
- ii. contacting the immobilized complex with at least one type of labeled nucleotide triphosphate (NTP), wherein each type of NTP is differently labeled with a detectable label which is released when said NTP is incorporated, and
- iii. detecting the incorporation of a labeled NTP into a single molecule of the primer by detecting a unique label released from the labeled NTP, to genotype or to sequence the target nucleic acid.
A method of genotyping or sequencing a target nucleic acid comprising the steps of:
- i. immobilizing onto a solid support a complex comprising a target nucleic acid and at least one nucleic acid polymerase;
- ii. contacting the immobilized complex with at least one type of labeled nucleotide triphosphate (NTP), wherein each type of NTP is differently labeled with a detectable label which is released when said NTP is incorporated, and iii. detecting the incorporation of a labeled NTP into a single nucleic acid that is complementary to the target nucleic acid by detecting a unique label released from the labeled NTP, to genotype or to sequence the target nucleic acid.
Dependencies are inferred from the claim text.