Multifunctional Verification Molecules for Oligonucleotide Synthesis Quality Control
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for synthesizing combinatorial libraries face inefficiencies due to limitations in assembling chemical subunits, reaction efficiency, and the challenge of accurately verifying and quantifying the synthesis of oligonucleotide probe molecules, often resulting in defective molecules and false results.
Innovation Solution
The development of multifunctional verification molecules with oligonucleotides containing coding regions and a chromatography agent, allowing for the identification and purification of accurately synthesized molecules through hybridization and PCR, enabling the separation of defective molecules and improving yield analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If a series of synthetic steps are used to form oligonucleotide probe molecules, then the probe molecules can be synthesized with desired functions, but the chance of defects increases with each synthetic step due to reaction failures and side reactions
Solution Approach 1:
The patent applies preliminary action by incorporating a verification oligonucleotide sequence into the probe molecule structure before synthesis is complete. This verification sequence serves as a built-in quality control mechanism that allows detection of synthesis defects at any stage, enabling early intervention and preventing propagation of defective molecules through subsequent synthesis steps.
Solution Approach 2:
The patent implements feedback by using the verification oligonucleotide sequence to monitor synthesis progress and detect defects. Through hybridization assays and sequencing, the verification sequence provides real-time information about synthesis accuracy, allowing researchers to identify and eliminate defective probe molecules from the library, thereby improving overall reliability.
2Productivity
If vast numbers of probe molecules are simultaneously tested for desired properties, then efficiency is preserved, but it becomes difficult to identify probe molecules with desired properties when libraries have sufficient diversity
Solution Approach 1:
The patent uses the verification oligonucleotide sequence as an intermediary between the probe molecule and the identification process. This verification sequence acts as a mediator that enables specific detection and identification of probe molecules with desired properties through hybridization assays, PCR amplification, and sequencing, allowing accurate identification even in large diverse libraries.
Solution Approach 2:
The patent applies copying by using PCR amplification to generate multiple copies of probe molecules that contain the verification oligonucleotide sequence. This amplification process creates sufficient copies for accurate sequencing and identification, enabling precise measurement of probe molecule properties while maintaining the ability to handle vast numbers of molecules in parallel.
3Ease of manufacture
If there is no easy or cost-effective method of determining if each reaction step was successful, then time and money are saved by avoiding analysis, but the result is a mixture of accurately formed and defective probe molecules
Solution Approach 1:
The patent extracts the verification function from complex analytical methods by incorporating a simple verification oligonucleotide sequence directly into the probe molecule. This extracted verification element can be detected through simple hybridization assays and sequencing, eliminating the need for expensive and time-consuming analysis of each reaction step while maintaining high synthesis verification capability.
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
This approach enhances the efficiency of synthesizing and verifying oligonucleotide probe molecules, reducing defects and false results by enabling cost-effective verification and purification, thereby improving the accuracy of combinatorial chemistry processes.
Implementation Method 1
G includes an oligonucleotide, the oligonucleotide comprising at least two coding regions, wherein the at least two coding regions are single stranded
Implementation Method 2
U is a chromatography agent
Data Source
AI summary
The present disclosure relates to multifunctional verification molecules, including molecules according to formula (I): G-L-(B)K-Q-U, wherein G, L, B, K, Q, and U are defined herein. The present disclosure also relates to methods of preparing and using such multifunctional verification molecules to remove defective multifunctional molecules and to quantify synthetic yield.