Competitive compositions of nucleic acid molecules for enrichment of rare-allele-bearing species

A composition, nucleic acid technology, applied in combinatorial chemistry, organic chemistry, sugar derivatives, etc.

Active Publication Date: 2017-02-22
RICE UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, to date, there is no accurate and inexpensive detection system that can be combined with multiple probe substances

Method used

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  • Competitive compositions of nucleic acid molecules for enrichment of rare-allele-bearing species
  • Competitive compositions of nucleic acid molecules for enrichment of rare-allele-bearing species
  • Competitive compositions of nucleic acid molecules for enrichment of rare-allele-bearing species

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0163] Experimental Results of Conditional Fluorescent Competitive Compositions

[0164] To experimentally verify the ability of the competing composition to enrich the target on the variants upon binding to the probe, experiments were designed and performed on conditionally fluorescent versions of the probe ( Figure 5 ). A probe is naturally dark until it hybridizes to a target or variant. By observing the fluorescent response to a sample of the variant with a small (loaded) amount of target, and the same amount of variant as in the absence of target, it can be deduced that enrichment is provided by the competing composition.

[0165] now refer to Figure 5 , 4 main products through image 3 The reactions described were formed; among these, 2 parameters involving the probe resulted in high fluorescence. The fluorescence generated by the target bound to the probe is denoted as f A , the fluorescence generated by the variant bound to the probe is denoted as f C , and the...

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Abstract

The present disclosure describes the thermodynamic design and concentrations necessary to design probe compositions with desired optimal specificity that enable enrichment, detection, quantitation, purification, imaging, and amplification of rare-allele-bearing species of nucleic acids (prevalence<1%) in a large stoichiometric excess of a dominant-allele-bearing species (wildtype). Being an enzyme-free and homogeneous nucleic acid enrichment composition, this technology is broadly compatible with nearly all nucleic acid-based biotechnology, including plate reader and fluorimeter readout of nucleic acids, microarrays, PCR and other enzymatic amplification reactions, fluorescence barcoding, nanoparticle-based purification and quantitation, and in situ hybridization imaging technologies.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Provisional Patent Application No. 61 / 981,588, filed April 18, 2014, which is incorporated herein by reference in its entirety. [0003] Statement of Government Rights [0004] This invention was made with government support under Grant No. EB015331 awarded by the National Institutes of Health. The government has certain rights in this invention. [0005] sequence listing [0006] This application contains a Sequence Listing electronically filed in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on April 15, 2015, is named 14-21011-WO_260947_00237_SL.txt and is 66,959 bytes in size. Background technique [0007] Nucleic acids serve as important molecular markers of disease, and the precise detection of specific DBA and RNA sequences forms the basis of many research and diagnostic products. However, many, if not most, prior art plat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C07H21/04C40B30/04
CPCC12Q1/6806C12Q2525/161C12Q2525/186C12Q2525/301C12Q2525/313C12Q2527/101C12Q2527/143C12Q2563/107C12Q2563/131C12Q1/6827
Inventor D·Y·张王珏皛
Owner RICE UNIV
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