| 摘要 |
Method for parallel sequencing analysis of nucleic acids (NA) in which single-stranded fragments (F), of 50-1000 bases, are generated, representing overlapping fragments of a complete sequence. (F) are then attached, as a complex (C) with one or primers, in random fashion to a reaction surface. Method for parallel sequencing analysis of nucleic acids (NA) in which single-stranded fragments (F), or 50-1000 bases, are generated, representing overlapping fragments of a complete sequence. (F) are then attached, as a complex (C) with one or primers, in random fashion to a reaction surface. The complementary strand of (F) is constructed using one or more polymerases in the following cycle: (a) incubating bound (C) with at least one polymerase and 1-4 modified nucleotides (N asterisk ), labeled with a fluorescent dye (FD), different for each N asterisk so that they can be distinguished, and N asterisk are modified at the 3'-end so that the polymerase, after incorporating one N asterisk into the growing complementary chain, can not incorporate another, but both FD and the terminating substance (TS) are cleavable; (b) extending the complementary strand by one N asterisk ; (c) washing to remove unincorporated N asterisk ; (d) detecting individual incorporated N asterisk from the characteristic fluorescence, with simultaneous detection of the relative positions of (C) on the surface; (e) removing FD and TS; (f) washing to eliminate FD and TS released, and optionally repeating the entire cycle. The relative positions of individual (C) and their sequences are determined through specific correlation of the signal detected in (d) in successive cycles. Independent claims are also included for: (a) a similar method for highly parallel analysis of gene expression using single-stranded gene products instead of F; (b) carriers for the new process; and (c) kits for the new process.
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