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Stoichiometric regulation of nucleic acid hybridization probes by helper oligonucleotide species

A nucleic acid probe and nucleic acid molecule hybridization technology, which is applied in the field of stoichiometric adjustment of nucleic acid hybridization probes by auxiliary oligonucleotide substances, and can solve the problems of high cost, inaccurate reaction energy, lack of precision and the like

Active Publication Date: 2021-06-11
RICE UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, reaction energy inaccuracies of up to 3 kcal / mol still occur due to imperfect biophysical models and literature parameters, especially in multiplexed environments
Empirical adjustments of operating conditions and probe sequences are often used to compensate for inaccuracies in probe design; however, these methods are expensive, time-consuming, and often lack precision

Method used

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  • Stoichiometric regulation of nucleic acid hybridization probes by helper oligonucleotide species
  • Stoichiometric regulation of nucleic acid hybridization probes by helper oligonucleotide species
  • Stoichiometric regulation of nucleic acid hybridization probes by helper oligonucleotide species

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Embodiment Construction

[0040] Protection implementation:

[0041] The protector implementation includes a probe nucleic acid molecule (denoted as C) and a helper nucleic acid molecule (denoted as protector or P), the probe nucleic acid molecule comprising first and second subsequences complementary to adjacent subsequences of the target, the The helper nucleic acid molecule comprises a third subsequence complementary to the first subsequence; P has a higher initial concentration than C. In some embodiments, C further comprises a fourth subsequence that is not complementary to the target sequence, and P further comprises a fifth subsequence that is complementary to the fourth subsequence. In some embodiments, each of C and P is an oligonucleotide, and the mixture of C and P is referred to as a toehold probe; Figure 1A An example of this implementation is shown in . More information on cohesive end probes can be found in WO 2015 / 094429A1, which is hereby incorporated by reference in its entirety.

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Abstract

The present invention describes methods for controlling the hybridization yield of nucleic acid probes by adjusting the relative concentrations of helper oligonucleotides relative to the probe and target. The helper oligonucleotide is partially or fully complementary to the probe or target and is released after the probe has hybridized to the target.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Provisional Patent Application No. 62 / 148,555, filed April 16, 2015, which is incorporated herein by reference in its entirety. [0003] Statement of Government Interests [0004] This invention was made with government support under Grant No. R00EB015331 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 8, 2016, is named 15-21010-WO_(260947.00262)_SL.txt and is 2,437 bytes in size. Background technique [0007] Nucleic acids encode vast amounts of biological and clinical information, and next-generation sequencing (NGS) is a promising class of methods to improve understanding of biology and inform medical decisions. Standard...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6811C12Q1/6832C12Q1/6876C07H21/02C07H21/04C07H1/06
CPCC07H21/02C07H21/04C07H1/06C12Q1/6811C12Q1/6832C12Q2525/161C12Q2525/204C12Q2527/143C12Q2535/131C12Q2537/1373C12Q2537/163C12Q2537/165C12Q2565/101C12Q2565/107C12Q1/6876
Inventor D·Y·张吴若嘉王珏皛
Owner RICE UNIV