In Vitro Nucleic Acid Isolation Using Oligonucleotide Tagged Microparticles
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Solution Overview
Problem
Current methods for isolating target nucleic acid sequences from a population are slow, expensive, and inefficient, requiring overnight bacterial transformation and colony growth, with limitations in accuracy and throughput.
Innovation Solution
The method involves attaching oligonucleotide tags to nucleic acid molecules, amplifying them, and using high-throughput sequencing and microparticles with complementary oligonucleotide sequences to isolate and identify target nucleic acid molecules through limiting dilution and amplification techniques.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional bacterial transformation and colony growth methods are used, then target nucleic acid sequences can be isolated, but the process is slow and time-consuming (overnight required)
Solution Approach 1:
The patent replaces the biological bacterial transformation system with an in vitro chemical system using oligonucleotide tags and microparticles. This substitution eliminates the need for overnight bacterial culture while maintaining isolation accuracy through controlled chemical hybridization and magnetic separation processes.
Solution Approach 2:
The patent performs preliminary tagging of nucleic acid molecules with oligonucleotide tags before the actual isolation process. This preliminary action allows for rapid subsequent separation using complementary tagged microparticles, eliminating the time-consuming bacterial transformation step while ensuring accurate target identification.
2Measurement precision
If traditional bacterial transformation methods are used, then target sequences can be identified, but the transformation is expensive
Solution Approach 1:
The patent uses disposable oligonucleotide tags and microparticles that can be synthesized cheaply and used in a single isolation cycle. These consumables replace expensive bacterial transformation reagents and antibiotic resistance markers, significantly reducing per-sample costs while maintaining high identification accuracy through sequence-specific hybridization.
Solution Approach 2:
The patent extracts only the essential isolation function from the complex bacterial transformation process, separating target sequences using simple oligonucleotide hybridization and magnetic bead separation. This extraction eliminates unnecessary expensive steps (bacterial culture, antibiotic selection) while preserving the core function of accurate sequence identification.
3Measurement precision
If traditional colony isolation and liquid media growth are used, then individual sequences can be identified, but the process is inefficient with low throughput
Solution Approach 1:
The patent merges multiple isolation steps into a single in vitro reaction mixture where tagged microparticles simultaneously capture multiple target sequences from the population. This consolidation allows parallel processing of numerous samples in one experiment, dramatically increasing throughput while maintaining precision through individual microparticle-sequence correspondence.
Solution Approach 2:
The patent transitions from two-dimensional colony growth on agar plates to three-dimensional manipulation of microparticles in suspension using magnetic fields. This dimensional change enables rapid mixing, separation, and processing of large numbers of samples simultaneously,大幅提升 throughput while maintaining precise target isolation through magnetic actuation.
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 significantly increases the accuracy, yield, and cost efficiency of nucleic acid synthesis and assembly, enabling the isolation of target sequences with high precision and speed, and the generation of large libraries with a high percentage of predetermined sequences.
Implementation Method 1
providing a plurality of microparticles, wherein each microparticle has an oligonucleotide sequence complementary to a portion of the nucleic acid molecules immobilized on its surface; forming a population of nucleic acid molecules hybridized to the complementary oligonucleotide sequence on the microparticles
Data Source
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AI summary
Methods and devices relate to the isolation of nucleic acids of interest from within a population of nucleic acids such as libraries of nucleic acid sequences.