초록 ▼

The present invention also relates to single molecule optical sequencing methods and systems for determining the nucleotide sequence of individual double stranded nucleic acid molecules elongated and fixed to a solid-surface by nicking the nucleic acid molecule, enzymatically adding labeled nucleoti

The present invention also relates to single molecule optical sequencing methods and systems for determining the nucleotide sequence of individual double stranded nucleic acid molecules elongated and fixed to a solid-surface by nicking the nucleic acid molecule, enzymatically adding labeled nucleotides and imaging the labeled nucleotides. The present invention also relates to methods and systems of single molecule optical sequencing using primer extension comprising the steps of elongating and fixing the nucleic acid molecule annealed with or to be annealed with at least one primer on a surface so that the nucleic acid remains accessible for enzymatic reactions with enzymes for the addition of labeled nucleotides; exposing the nucleic acid molecule annealed with at least one primer to a polymerase and dideoxy nucleotide; exposing the nucleic acid molecule to a polymerase and nucleotides including labeled nucleotides to produce a primer extension nucleic acid molecule; and imaging the labeled primer extension molecule to produce an image. The invention further relates to methods of imaging single or multiple labeled nucleotides enzymatically added to nicked individual double stranded nucleic acid molecules. The invention also provides a method of analysis of the images of labeled nucleotides or primer extension products using Bayesian estimation to determine the sequence of the nucleic acid molecules.

대표청구항 ▼

[ What is claimed is:] [1.]1. A method for imaging a single labeled nucleotide on an individual double stranded nucleic acid molecule, comprising:(a) nicking a double stranded nucleic acid molecule elongated and fixed onto a surface so that the double stranded nucleic acid molecule remains accessibl

[ What is claimed is:] [1.]1. A method for imaging a single labeled nucleotide on an individual double stranded nucleic acid molecule, comprising:(a) nicking a double stranded nucleic acid molecule elongated and fixed onto a surface so that the double stranded nucleic acid molecule remains accessible for enzymatic reactions with enzymes for the addition of a labeled nucleotide creating a nicked strand;(b) enzymatically adding a single nucleotide comprising a label; and(c) imaging the added label.