Nanopore Sequencing for Real-Time Nucleic Acid Isolation
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Solution Overview
Problem
Current methods for isolating accurate nucleic acid strands from mixed libraries are time-consuming and costly, particularly in nucleic acid sequencing and synthesis applications, where errors in length and sequence identity are common, necessitating rapid and cost-efficient techniques for separating desired strands.
Innovation Solution
The use of nanopore sequencing devices and substrate-based sequencing technologies, which involve localized sequencing and selective release of desired nucleic acid strands based on sequence, length, and methylation status, utilizing photocleavable linkers and controlled voltage to isolate accurate strands from inaccurate ones.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If molecular cloning followed by sequencing is used to isolate accurate nucleic acid strands, then the isolation accuracy is improved, but the time required increases to one to three weeks and the cost increases considerably
Solution Approach 1:
The patent applies preliminary action by performing sequencing and characterization of nucleic acid strands during the synthesis process itself, rather than after completion. The system sequences individual strands as they are synthesized and makes real-time determination of accuracy, allowing immediate separation of accurate strands from inaccurate ones, thereby eliminating the need for subsequent cloning and sequencing steps that would take one to three weeks.
Solution Approach 2:
The patent replaces the mechanical biological process of molecular cloning with an automated electrochemical system. Instead of using biological replication and physical separation methods, the system uses nanopore sequencing technology combined with automated voltage control and computer processing to identify and separate accurate nucleic acid strands, significantly reducing time and cost while maintaining high accuracy.
2Measurement precision
If molecular cloning followed by sequencing is used to isolate accurate nucleic acid strands, then the isolation accuracy is improved, but the cost increases considerably
Solution Approach 1:
The patent applies preliminary action by performing sequencing and characterization of nucleic acid strands during the synthesis process itself, rather than after completion. The system sequences individual strands as they are synthesized and makes real-time determination of accuracy, allowing immediate separation of accurate strands from inaccurate ones, thereby eliminating the need for subsequent cloning and sequencing steps that would take one to three weeks.
Solution Approach 2:
The patent replaces the mechanical biological process of molecular cloning with an automated electrochemical system. Instead of using biological replication and physical separation methods, the system uses nanopore sequencing technology combined with automated voltage control and computer processing to identify and separate accurate nucleic acid strands, significantly reducing time and cost while maintaining high accuracy.
3Measurement precision
If conventional sequencing methods are used, then the sequence determination is achieved, but the process is not rapid enough for efficient nucleic acid synthesis
Solution Approach 1:
The patent applies preliminary action by performing sequencing and characterization of nucleic acid strands during the synthesis process itself, rather than after completion. The system sequences individual strands as they are synthesized and makes real-time determination of accuracy, allowing immediate separation of accurate strands from inaccurate ones, thereby eliminating the need for subsequent cloning and sequencing steps that would take one to three weeks.
Solution Approach 2:
The patent implements continuity of useful action by maintaining continuous sequencing and evaluation of nucleic acid strands throughout the synthesis process. The system operates continuously to sequence individual strands, evaluate their accuracy, and separate accurate strands from inaccurate ones in real-time, eliminating idle time and sequential processing steps, thereby dramatically increasing productivity.
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
Enables rapid and cost-effective isolation of accurate nucleic acid strands by determining sequence and properties in real-time, allowing for efficient separation and collection of desired strands, reducing the need for extensive cloning and sequencing processes.
Implementation Method 1
inducing a flow of current through each nanopore, such that individual nucleic acid strands enter into the nanopores housed within the membrane
Implementation Method 2
A difference in electrical potential is generated between the two chambers, such that nucleic acid strands begin the process of translocating through the nanopore
Implementation Method 3
utilizing photocleavable linkers and controlled voltage to isolate accurate strands from inaccurate ones
Data Source
AI summary
The present disclosure provides systems and methods for isolation of nucleic acids. In particular, provided herein are nanopore-based or substrate-based sequencing systems and methods of use thereof for isolation of desired nucleic acids from a mixed library containing both accurate and inaccurate strands.