Probe/mobility modifier complexes for multiplex nucleic acid detection

• US 20050112594A1
• Filed: 01/29/2004
• Published: 05/26/2005
• Est. Priority Date: 03/15/1999
• Status: Abandoned Application

• Current Assignee: Applera Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applera Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Grossman, Paul D.
• Application Number:
  US10/769,288
• Publication Number:
  US 20050112594A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 1/6827 for detection of mutation or polymorphism
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 1/6858 Allele-specific amplification
• CPC Class Codes: C12Q 2525/161 incorporating target specific and non-target specific sites
• CPC Class Codes: C12Q 2535/125 Allele specific primer extension
• CPC Class Codes: C12Q 2537/143 Multiplexing, i.e. use of multiple primers or probes in a single reaction, usually for simultaneously analyse of multiple analysis
• CPC Class Codes: C12Q 2563/179 the label being a nucleic acid

Abstract

Compositions and methods for the analysis of multiple nucleic acid target sequences are disclosed. The compositions comprise a probe comprising a target-specific portion for sequence-specific hybridization to a target nucleic acid sequence, and a tag; and a mobility-modifier comprising a tail and a tag complement for binding to the tag. The associated methods generally comprise the steps of providing a sample potentially containing one or more target nucleic acid sequences; providing one or more probes, each probe comprising a target-specific portion and a tag; providing one or more mobility modifiers, each mobility modifier comprising a tag complement and a tail; contacting the probe(s) and the target nucleic acid sequence(s) under conditions effective for sequence-dependent hybridization of the probe(s) and the target nucleic acid sequence(s); contacting the probe(s) and the mobility-modifier(s) under conditions suitable for selectively binding the probe(s) to the mobility modifier(s), thereby forming one or more a probe/mobility modifier complex(s); and analyzing the probe/mobility modifier complex(s) using a mobility-dependent analysis technique.

Claims

1. A binary composition for detecting one or more target nucleic acid sequences comprising:

a probe comprising a target-specific portion for sequence-specific hybridization to a target nucleic acid sequence, and a tag; and

a mobility-modifier comprising a tail and a tag complement for binding to the tag.

2. The composition of claim 1 wherein the target-specific portion comprises polynucleotide.

3. The composition of claim 2 wherein the target-specific portion comprises a 3′

-hydroxyl group.

4. The composition of claim 2 wherein the target-specific portion comprises PNA.

5. The composition of claim 1 wherein the tag portion comprises polynucleotide.

6. The composition of claim 5 wherein the tag portion is PNA.

7. The composition of claim 1 wherein both the tag and tag complement are polynucleotide, and one of the tag complement and tag comprises a sequence selected from the group consisting of (CAG)_n and (TCC)_n wherein n is 1 to 10.

8. The composition of claim 1 wherein the mobility modifier comprises a tail portion.

9. The composition of claim 8 wherein the tail is a polymer.

10. The composition of claim 9 wherein the polymer is selected from the group consisting of polyethyleneoxide and polypeptide.

11. The composition of claim 1 further comprising a hybridization enhancer.

6.1. 1 (canceled)

12-25. -25. (canceled)