Solid-phase clonal amplification and related methods

摘要

The present invention provides methods and compositions for analyzing nucleic acid sequences. In some aspects, the methods utilize clonal objects, such as DNA balls, that have been captured on beads. Using the methods described here, compositions are fabricated wherein a bead and one clonal object are affinity bound or hybridized to each other through an affinity binding patch or hybridization patch on the surface of the bead. The invention also provides a population of beads having affinity bound or hybridized clonal objects at a ratio of 1:1. The invention additionally provides methods for amplifying a target nucleic acid molecule utilizing the compositions described herein.

主权项

1. A method of fabricating a bead having one clonal object comprising:
(a) providing a bead comprising a plurality of first polynucleotides; (b) providing a solid surface comprising a plurality of second polynucleotides patterned into patches on said surface, wherein said second polynucleotides each comprise a cleavable moiety, wherein one clonal object is hybridized to one polynucleotide patch on said surface, and wherein said one clonal object comprises a single tandemly repeated target nucleic acid molecule and a plurality of binding sites; (c) hybridizing said first polynucleotides to said plurality of binding sites on said clonal object, thereby forming multiple hybridization events between said clonal object and an immobilized bead on the solid surface, and (d) cleaving said second polynucleotides at said cleavable moiety so as to retain said clonal object, thereby fabricating a bead having one clonal object.