초록 ▼

The invention provides a method for sequencing each polynucleotide of a population by using an oligonucleotide tag assigned to each such polynucleotide for transfering sequence information to a tag complement located on a spatially addressable array of such complements. That is, a unique tag is atta

The invention provides a method for sequencing each polynucleotide of a population by using an oligonucleotide tag assigned to each such polynucleotide for transfering sequence information to a tag complement located on a spatially addressable array of such complements. That is, a unique tag is attached to each polynucleotide of a population which can be copied and used to shuttle sequence information to its complement at a fixed position on an array of such complements. After a tag hybridizes with its complement, a signal is generated that is indicative of the transferred sequence information. Sequences of the tagged polynucleotides are determined by repeated cycles of information transfer and signal detection at the positions of the corresponding tag complements.

대표청구항 ▼

[ I claim:] [1.] A method for simultaneously determining the nucleotide sequences of a population of polynucleotides, the method comprising the steps of:(a) attaching an oligonucleotide tag from a repertoire of tags to each polynucleotide of the population to form tag-polynucleotide conjugates such

[ I claim:] [1.] A method for simultaneously determining the nucleotide sequences of a population of polynucleotides, the method comprising the steps of:(a) attaching an oligonucleotide tag from a repertoire of tags to each polynucleotide of the population to form tag-polynucleotide conjugates such that substantially all different polynucleotides have different oligonucleotide tags attached;(b) selectively amplifying tag-polynucleotide conjugates with primers whose 3' ends form perfectly matched duplexes with one or more terminal nucleotides of the polynucleotides of the population;(c) labeling each tag of the selectively amplified tag-polynucleotide conjugates according to the identity of the one or more terminal nucleotides of its associated polynucleotide;(d) cleaving the tags from the selectively amplified tag-polynucleotide conjugates; and(e) sorting the labeled tags onto a spatially addressable array of tag complements for detection of the labeled tags and identification of the one or more nucleotides of each polynucleotide.