IP Library › Granted Patent US 8,685,899
Granted Patent B2
US 8,685,899 · App. 12/031,165 · Granted Apr 1, 2014

Methods, reagents and kits for detection of nucleic acid molecules

Inventors: Robert C. Getts (Collegeville, PA); James Kadushin (Gilbertsville, PA); Jessica Bowers (Harleysville, PA)
Assignee: Genisphere Inc.
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Quick Facts
Patent No.
US 8,685,899
App. No.
12/031,165
Granted
Apr 1, 2014
Kind
B2
Abstract

Methods, reagents and kits are provided for the production and use in detection assays of labeled nucleic acid molecules wherein a labeling molecule is attached directly to the 3′ end of the nucleic acid molecules.

Claims (10)

1. A multi-labeled polymeric scaffold to which a plurality of label molecules capable of emitting or producing a detectable signal is attached, wherein the multi-labeled polymeric scaffold comprises an oligonucleotide extension sequence comprising a 5′ phosphate group which is capable of hybridization bonding to a nucleic acid sequence, wherein the multi-labeled polymeric scaffold is a linear dendritic polynucleotide composition having a plurality of single stranded regions to which one or more labeled oligonucleotides are hybridized, said linear dendritic polynucleotide composition comprising first, second and third polynucleotide monomers bonded together by hybridization in a 5′-3′ orientation, each polynucleotide monomer, prior to being hybridization bonded to one another, having first, second and third single stranded hybridization regions; and in said linear dendritic polynucleotide composition the third single stranded hybridization region of the first polynucleotide monomer being hybridization bonded to the first single stranded hybridization region of the second polynucleotide monomer, and the third single stranded hybridization region of the second polynucleotide monomer being hybridization bonded to the first single stranded hybridization region of the third polynucleotide monomer, and the first single strand region of the first polynucleotide monomer is capable of hybridization bonding to the nucleic acid sequence,

wherein the second single stranded hybridization regions within said linear dendritic polynucleotide composition are hybridization bonded to one or more labeled oligonucleotides comprising one or more label molecules and wherein the multi-labeled polymeric scaffold has a total molecular weight of about 50 to about 350 kDa.

2. The multi-labeled polymeric scaffold of claim 1 , wherein the nucleic acid sequence is a bridging oligonucleotide, and wherein the extension sequence is hybridization bonded to said bridging oligonucleotide.

3. The multi-labeled polymeric scaffold of claim 2 , wherein the bridging oligonucleotide is capable of hybridization bonding to a nucleic acid molecule separate and distinct from the polymeric scaffold.

4. The multi-labeled polymeric scaffold of claim 3 , wherein the nucleic acid molecule separate and distinct from the polymeric scaffold is a poly(A) tailed RNA molecule.

5. The multi-labeled polymeric scaffold of claim 1 , wherein the label molecules are fluorophores.

6. The multi-labeled polymeric scaffold of claim 1 , wherein the scaffold is crosslinked following assembly.

7. The multi-labeled polymeric scaffold of claim 1 , wherein the label molecules are spaced at intervals of at least 10 nucleotides.

8. The multi-labeled polymeric scaffold of claim 7 , wherein the scaffold has a molecular weight of 165 kDa and comprises 15 label molecules.

9. The multi-labeled polymeric scaffold of claim 1 , wherein the third single stranded hybridization region of said third polynucleotide monomer is hybridization bonded to one or more labeled oligonucleotides comprising one or more label molecules.

Assignments (5)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jan 7, 2010
From: GENISPHERE INC.
To: CF PARTNERS ACQUISITION III, LLC
Reel/Frame 023741/0896 →
CHANGE OF NAME Recorded Jan 7, 2010
From: CF PARTNERS ACQUISITION III, LLC
To: GENISPHERE, LLC
Reel/Frame 023741/0907 →
CHANGE OF OWNERSHIP Recorded Sep 30, 2009
From: DATASCOPE INVESTMENT CORP.
To: GENISPHERE INC.
Reel/Frame 023302/0317 →
CORRECTIVE ASSIGNMENT TO CORRECT THE NAME OF ASSIGNEE PREVIOUSLY RECORDED ON REEL 021706 FRAME 0090. ASSIGNOR(S) HEREBY CONFIRMS THE NAME OF ASSIGNEE AS DATASCOPE INVESTMENT CORP. Recorded Jan 5, 2009
From: GETTS, ROBERT C.; KADUSHIN, JAMES; BOWERS, JESSICA
To: DATASCOPE INVESTMENT CORP.
Reel/Frame 022053/0817 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 20, 2008
From: GETTS, ROBERT C.; KADUSHIN, JAMES; BOWERS, JESSICA
To: DATASCOPE INVESTGMENT CORP.
Reel/Frame 021706/0090 →
Continuity (2)
Provisional Application 60901361 · Feb 14, 2007
Related Publication 20080300142A1 · Dec 4, 2008