Modified Nucleic Acid Units for Single-Base Library Purification
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Solution Overview
Problem
Existing nucleic acid purification methods struggle to efficiently purify error-free nucleic acids from libraries with varying types and sequences, leading to low throughput and recovery efficiency.
Innovation Solution
A method involving the use of modified nucleic acid units with terminator moieties to selectively purify nucleic acids of desired lengths by binding complementary nucleic acid chains, utilizing next-generation sequencing instruments for high-throughput purification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional PAGE or HPLC purification methods are used, then nucleic acids of intended length can be selected, but purification efficiency is low when various types of molecules are in a nucleic acid library
Solution Approach 1:
The invention changes the chemical parameter of nucleic acid units by introducing modified nucleic acid units with terminator moieties during binding. This allows selective purification based on the presence/absence of the terminator at specific positions, enabling high-throughput purification while maintaining precision across diverse nucleic acid sequences in the library
Solution Approach 2:
The modified nucleic acid unit with terminator moiety acts as an intermediary that enables selective binding and purification. The terminator serves as a chemical marker that facilitates the selection of error-free nucleic acids without requiring individual sequence analysis, thereby improving both precision and efficiency simultaneously
2Manufacturing precision
If error-correction enzyme method is used, then error-free nucleic acids can be purified, but it is difficult to apply when various types of molecules are in the nucleic acid library
Solution Approach 1:
The modified nucleic acid unit with terminator moiety provides a universal purification mechanism that works across all types of nucleic acid sequences in the library. Unlike error-correction enzymes that require specific recognition sequences, the terminator-based method universally marks error-free nucleic acids regardless of their sequence composition, thereby improving adaptability while maintaining precision
3Manufacturing precision
If NGS sequence analysis method is used, then error-free nucleic acids can be identified, but recovery efficiency is low because one type of nucleic acid must be recovered individually
Solution Approach 1:
The invention performs preliminary chemical modification by incorporating terminator-containing modified nucleic acid units during the binding process. This preliminary action marks error-free nucleic acids with a detectable terminator signal before purification, enabling simultaneous identification and recovery of multiple nucleic acid types in high throughput, thereby improving both accuracy and efficiency
4Productivity
If conventional purification methods are used, then nucleic acids can be purified, but there is no technology capable of purifying nucleic acids at high throughput regardless of the complexity of the library
Solution Approach 1:
The invention changes the chemical parameter of nucleic acid units by introducing modified nucleic acid units with terminator moieties during binding. This allows selective purification based on the presence/absence of the terminator at specific positions, enabling high-throughput purification while maintaining precision across diverse nucleic acid sequences in the library
Solution Approach 2:
The invention replaces mechanical separation methods (PAGE, HPLC) with a chemical recognition system using terminator-containing modified nucleic acid units. This substitution enables automated, high-throughput purification while maintaining single-base resolution precision, as the chemical terminator provides a clear molecular signal for selection
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 high-throughput purification of error-free nucleic acids with single-base resolution, regardless of sequence or complexity, significantly improving purification efficiency.
Implementation Method 1
obtaining a library of complementary nucleic acids by binding complementary nucleic acid units to each base of the strand of the template nucleic acids
Data Source
AI summary
Provided are a method for purifying a nucleic acid library, and a kit, the method comprising the steps of: providing a nucleic acid library comprising single-stranded template nucleic acids; obtaining a library of complementary nucleic acids by binding complementary nucleic acid units to each base of the strand of the template nucleic acids; introducing at least one modified nucleic acid unit during the binding process of the nucleic acid units; and selectively selecting a nucleic acid having a desired length from the library of complementary nucleic acids using the modified nucleic acid unit. According to the present invention, the nucleic acid library may be purified regardless of the complexity, sequence or length of the nucleic acid library, and nucleic acids having different lengths may be simultaneously purified. The purification may be carried out through direct experiment or using a next-generation sequencing instrument.