Methods and compositions for tagging and identifying polynucleotides

摘要

The invention provides methods and compositions for attaching oligonucleotide tags to polynucleotides for the purpose of carrying out analytical assays in parallel and for decoding the oligonucleotide tags of polynucleotides selected in such assays. Words, or subunits, of oligonucleotide tags index submixtures in successively more complex sets of submixtures (referred to herein as “tiers” of submixtures) that a polynucleotide goes through while successive words are added to a growing tag. By identifying each word of an oligonucleotide tag, a series of submixtures is identified including the first submixture that contains only a single polynucleotide, thereby providing the identity of the selected polynucleotide. The analysis of the words of an oligonucleotide tag can be carried out in parallel, e.g. by specific hybridization of the oligonucleotide tag to its tag complement on an addressable array; or such analysis can be carried out serially by successive specific hybridizations of labeled word complements, or the like.

主权项

1. A method of sequencing comprising:
sequencing a plurality of cDNAs, wherein the cDNAs are obtained from a plurality of different sources, and:
(a) each of the cDNAs comprises: (i) a tag that indicates from which source a cDNA is derived and (ii) separate from the tag, a binding site for a sequencing primer;(b) the sequencing is done using the sequencing primer; and(c) the cDNAs have been selected from a pooled cDNA sample; to obtain, for each of the sequenced cDNAs, the nucleotide sequence of the tag associated with the cDNA and the nucleotide sequence of at least a portion of a cDNA.