Size-Selected Nucleic Acid Library Preparation via Affinity Binding
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Solution Overview
Problem
Current methods for size selection of nucleic acid libraries are expensive, time-consuming, and inefficient, particularly for isolating long DNA strands, as they require large DNA quantities and are prone to contamination with shorter DNA strands.
Innovation Solution
The method involves contacting a nucleic acid sample with a nucleic acid binding reagent containing an affinity tag, such as desthiobiotin, under specific conditions to selectively bind shorter nucleic acids, allowing longer nucleic acids to be separated and purified, thus producing a size-selected nucleic acid library.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional size selection methods (e.g., BluePippin system) are used, then nucleic acid size selection can be achieved, but the process becomes expensive, time-consuming, and requires large DNA quantities
Solution Approach 1:
The patent extracts and removes shorter nucleic acid strands from the library using a nucleic acid binding reagent with an affinity tag that specifically binds to nucleic acids of less than the desired length. This selective extraction allows longer nucleic acids to be separated and purified, directly addressing the size selection precision requirement while improving throughput by eliminating the need for conventional time-consuming systems
Solution Approach 2:
The patent introduces a nucleic acid binding reagent as an intermediary substance that mediates the size selection process. This reagent, containing an affinity tag, selectively interacts with shorter nucleic acid strands, enabling their removal from the library without requiring complex conventional size selection equipment, thus improving both precision and productivity
2Manufacturing precision
If conventional size selection methods are used, then nucleic acid size selection can be achieved, but the process becomes expensive and requires large microgram quantities of DNA
Solution Approach 1:
The patent extracts and removes shorter nucleic acid strands from the library using a nucleic acid binding reagent with an affinity tag that specifically binds to nucleic acids of less than the desired length. This selective extraction allows longer nucleic acids to be separated and purified, directly addressing the size selection precision requirement while improving throughput by eliminating the need for conventional time-consuming systems
Solution Approach 2:
The patent changes the binding parameters of the nucleic acid binding reagent, including the duration of contacting and the concentration of the reagent, to optimize selective binding of shorter strands. By adjusting these parameters, the method achieves effective size selection with reduced DNA quantities compared to conventional methods requiring large microgram amounts
3Manufacturing precision
If conventional size selection methods are used, then nucleic acid size selection can be achieved, but the process becomes time-consuming
Solution Approach 1:
The patent extracts and removes shorter nucleic acid strands from the library using a nucleic acid binding reagent with an affinity tag that specifically binds to nucleic acids of less than the desired length. This selective extraction allows longer nucleic acids to be separated and purified, directly addressing the size selection precision requirement while improving throughput by eliminating the need for conventional time-consuming systems
Solution Approach 2:
The patent performs preliminary binding of the nucleic acid binding reagent to shorter strands before separation. By pre-conditioning the reagent and optimizing binding conditions (duration and concentration), the method enables rapid and efficient size selection, reducing the overall time required compared to conventional sequential approaches
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 enables efficient and cost-effective size selection of nucleic acid libraries, specifically targeting long DNA strands while minimizing contamination from shorter strands, thereby enhancing the throughput of long read lengths in sequencing applications.
Implementation Method 1
contacting a nucleic acid sample and a nucleic acid binding reagent including an affinity tag, under conditions in which nucleic acids of less than a desired length are substantially bound to the nucleic acid binding reagent
Data Source
AI summary
Provided are methods of producing size-selected nucleic acid libraries. The methods include contacting a nucleic acid sample and a nucleic acid binding reagent including an affinity tag, under conditions in which nucleic acids of less than a desired length are substantially bound to the nucleic acid binding reagent and nucleic acids of the desired length are substantially not bound to the nucleic acid binding reagent. The conditions include the duration of the contacting, the concentration of the nucleic acid binding reagent, or both. The methods further include separating, using the affinity tag, the nucleic acids of less than the desired length bound to the nucleic acid binding reagent from the nucleic acids of the desired length not bound to the nucleic acid binding reagent, to produce a size-selected nucleic acid library. Compositions and kits that find use, e.g., in practicing the methods of the present disclosure, are also provided.