Miniaturized, high-throughput nucleic acid analysis

摘要

The present invention is directed to method for analyzing multiple nucleic acid molecules of interest comprising in the steps of (i) providing a plurality of beads, characterized in that each bead comprises at least two sequence specific amplification primers, further characterized in that at least one of the primers is bound to the bead via a cleavable linker, (ii) capturing the nucleic acid molecules of interest from a sample, (iii) clonally isolating the plurality of beads, (iv) cleaving the at least one primer, (v) clonally amplifying the nucleic acid thereby creating multiple amplification products, and (vi) analyzing the amplification products.

主权项

1. A method for analyzing a sample comprising nucleic acid molecules of interest, the method comprising the steps of:
(a) providing a plurality of beads and a plurality of sequence specific amplification primer pairs, wherein each bead comprises one pair of sequence specific amplification primers comprising a first primer and a second primer, and wherein the first primer is bound to the bead via photo-cleavable linker, (b) capturing onto the beads the nucleic acid molecules of interest from the sample by hybridization of the nucleic acid molecules to the second primer, (c) clonally isolating said plurality of beads with captured nucleic acid molecules, (d) cleaving the first primer by photo-induction, while the second primer remains covalently attached to the bead, (e) clonally amplifying said nucleic acid, thereby generating multiple amplification products, and (f) analyzing said amplification products.