Hybridization Buffer with Blocking Nucleic Acid for Specific Detection

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

The presence of non-target nucleic acids decreases the efficiency of hybridization and detection of target nucleic acids with nucleic acid probes, leading to non-specific hybridization and reduced detection accuracy.

Innovation Solution

A buffer composition for hybridization that includes a blocking nucleic acid with a nucleotide sequence complementary to non-target nucleic acids, which is designed to suppress non-specific hybridization by hybridizing with the non-target nucleic acids, allowing specific hybridization of the target nucleic acids with the probe nucleic acids to occur.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional blocking agents (nucleic acid components, surfactants, proteins) are used to suppress non-specific hybridization, then non-specific hybridization is partially suppressed, but blocking effects are insufficient when non-target nucleic acids are abundantly present

Engineering Contradiction:

Improveblocking effectVSAvoiddetection accuracy

Core Design Contradiction:

ReliabilityVSMeasurement precision

Solution Approach 1:

The patent introduces a blocker probe as an intermediary molecule that specifically binds to non-target nucleic acids, preventing them from interacting with the capture sequence probe. This mediator resolves the contradiction by providing targeted blocking action that conventional agents cannot achieve, thereby improving both blocking effectiveness and detection accuracy simultaneously

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The blocker probe is designed with specific local properties (complementary nucleotide sequence matching the non-target nucleic acid) rather than using general-purpose blocking agents. This localized specificity allows the blocking agent to precisely target only the problematic non-target sequences without affecting overall hybridization efficiency, thus improving both reliability and measurement precision

Inventive Principle:

Principle #3Local quality

2Measurement precision

If blocking agents are added to suppress non-specific hybridization, then detection specificity is improved, but detection efficiency decreases due to presence of non-target nucleic acids

Engineering Contradiction:

Improvedetection specificityVSAvoiddetection efficiency

Core Design Contradiction:

Measurement precisionVSProductivity

Solution Approach 1:

The blocker probe is added to the reaction solution before the hybridization step, allowing it to pre-bind to non-target nucleic acids and form stable complexes. This preliminary action prevents non-target sequences from interfering with subsequent probe hybridization, thereby maintaining high detection efficiency while achieving specific detection of target nucleic acids

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent converts the harmful presence of abundant non-target nucleic acids into a beneficial situation by using them as binding targets for the blocker probe. The non-target sequences that would otherwise cause non-specific hybridization and reduce detection efficiency are now specifically sequestered by the blocker probe, allowing efficient and specific detection of target nucleic acids to proceed

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The use of the buffer composition effectively suppresses non-specific hybridization, enhancing the detection efficiency and accuracy of target nucleic acids even in the presence of non-target nucleic acids, particularly when used in microarray systems.

Implementation Method 1

hybridization means a phenomenon in which nucleic acids hydrogen-bond to each other through complementary base pairing

Methodology Applied

Scientific EffectHybridization: Chemical Bonding

Implementation Method 2

nucleic acids hydrogen-bond to each other through complementary base pairing

Methodology Applied

Scientific EffectHydrogen bonding: Chemical Bonding

Implementation Method 3

a method involving properly regulating the salt concentration and reaction temperature of a reaction solution

Methodology Applied

Scientific EffectTemperature control: Thermal Radiation

Data Source

PatentUS10450601B2Buffer composition for hybridization use, and hybridization method

Publication Date: 2019.10.22 TOYO KOHAN CO LTD

AI summary

A buffer composition for hybridization of a target nucleic acid is provided. The nucleic acid can include a nucleotide to be detected with a nucleic acid probe. The probe can contain a nucleotide sequence complementary to the target nucleic acid. The buffer can include a blocking nucleic acid having a nucleotide sequence complementary to a non-target nucleic acid having a nucleotide not to be detected corresponding to the nucleotide to be detected. The buffer composition can suppress non-specific hybridization to the nucleic acid probe even when a non-target nucleic acid is present. The use of the buffer composition can achieve excellent detection efficiency of the target nucleic acid.