RNA Library Construction Using Guide Nucleic Acid Mediator

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Solution Overview

Problem

Existing methods for constructing RNA libraries are inefficient and time-consuming, particularly in the ligation of adapter nucleic acid molecules, which hampers the rapid and accurate construction of RNA libraries for disease-related genetic information.

Innovation Solution

A method involving the ligation of a first adapter nucleic acid molecule to the 3' end of a target RNA, followed by complementary strand polymerization to synthesize cDNA, removal of RNA, ligation of a second adapter nucleic acid molecule with the aid of a guide nucleic acid molecule, and subsequent PCR amplification to enhance the construction efficiency of the RNA library.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If conventional ligation methods are used to construct RNA libraries, then the library construction can be completed, but the ligation efficiency of adapter nucleic acid molecules is low and the construction time is long

Engineering Contradiction:

Improvelibrary construction efficiencyVSAvoidconstruction time

Core Design Contradiction:

ProductivityVSLoss of time

Solution Approach 1:

A guide nucleic acid molecule is introduced as an intermediary to facilitate the ligation between the second adapter nucleic acid molecule and the cDNA. The guide molecule hybridizes to both the cDNA and the adapter, forming a stable complex that significantly enhances ligation efficiency and reduces construction time

Inventive Principle:

Principle #24Intermediary (Mediator)

2Manufacturing precision

If multiple ligation steps are performed sequentially, then the RNA library can be constructed with proper adapter integration, but the overall construction process becomes time-consuming

Engineering Contradiction:

Improveadapter ligation accuracyVSAvoidconstruction throughput

Core Design Contradiction:

Manufacturing precisionVSProductivity

Solution Approach 1:

The guide nucleic acid molecule is designed and prepared in advance to contain hybridization regions for both the cDNA and the second adapter. This preliminary preparation enables the guide molecule to mediate the ligation process efficiently, ensuring accurate adapter integration while reducing the time required for multiple sequential steps

Inventive Principle:

Principle #10Preliminary action

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 proposed method significantly improves the construction efficiency and reduces the time required for RNA library construction while maintaining accuracy, by enhancing the ligation efficiency of adapter nucleic acid molecules through the use of a guide nucleic acid molecule.

Implementation Method 1

by adding a guide nucleic acid molecule, the ligation efficiency of the adapter nucleic acid molecules can be increased

Methodology Applied

Scientific EffectHybridization:

Implementation Method 2

synthesizing cDNA through complementary strand polymerization of the molecule ligated in the step a)

Methodology Applied

Scientific EffectReverse transcription:

Data Source

PatentEP4549580A1RNA sequencing library construction method

Publication Date: 2025.05.07 XENOHELIX CO LTD

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AI summary

The present invention relates to a method for constructing an RNA library, which includes ligating of a first adapter nucleic acid molecule and a second adapter nucleic acid molecule. Additionally, the method of the present invention, by including a guide nucleic acid molecule, can increase the ligation efficiency of the adapter nucleic acid molecules, thereby increasing the efficiency of RNA library construction.