Solid support for high-throughput nucleic acid analysis

• US 9,347,092 B2
• Filed: 02/15/2010
• Issued: 05/24/2016
• Est. Priority Date: 02/25/2009
• Status: Active Grant

• Current Assignee: Roche Molecular Systems (Roche Holding AG)
• Original Assignee: Roche Molecular Systems (Roche Holding AG)
• Inventors: Roesler, Angelika, Froehlich, Thomas, Heindl, Dieter
• Primary Examiner(s): Gross, Christopher M
• Application Number:
  US12/705,679
• Publication Number:
  US 20100248991A1
• Time in Patent Office: 2,290 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: C12Q 1/6844 Nucleic acid amplification reactions
• CPC Class Codes: C12Q 2523/319 Photocleavage, photolysis, photoactivation
• CPC Class Codes: C12Q 2525/197 incorporating a spacer/coupling moiety
• CPC Class Codes: C12Q 2563/149 Particles, e.g. beads
• CPC Class Codes: C12Q 2565/537 characterised by the capture oligonucleotide acting as a primer

Abstract

The present invention provides a solid support which is preferably a bead comprising at least two sequence specific amplification primers wherein at least one primer is bound to the support with an inducible cleavable linker. The present invention also provides various method for preparing a solid support comprising at least two sequence specific primers, further characterized in that at least one of the primers is cleavable.

Claims

1. An emulsion comprising two or more types of micelle, each micelle comprising:

a bead, said bead comprising at least one pair of sequence specific amplification primers specific for a target nucleic acid wherein the at least one pair includes a first primer and a second primer, said first primer comprising a first nucleotide sequence and said second primer comprising a second nucleotide sequence different from the first nucleotide sequence,

said first primer bound to said bead with an inducible photo-cleavable linker via a functional moiety selected from the group consisting of carboxy, aldehyde, azide, alkyne, amino, thiol, sulfonylalken, iodoacetyl, amino hydrazine, hydroxylamino, and maleimido, and

said second primer is covalently attached to the bead via a second linker different from the inducible photo-cleavable linker of the first primer and is not cleavable under cleavage conditions for the first primer, and

wherein the at least one pair of sequence specific amplification primers in each of the two or more types of micelle is specific for a different target nucleic acid.

2. The emulsion of claim 1, wherein said bead is comprised of a material selected from the group consisting of silicon, titanium dioxide, aluminum oxide, lanthanide oxide, glass, silicates polystyrene, cellulose, Sepharose, and polyamide.

3. The emulsion of claim 1, wherein the first primer carries a detectable tag.

4. The emulsion of claim 1, wherein said bead has a size between about 10 μ

m to about 250 μ

m.