Solid-Support Nucleic Acid Loading with Poloxamer Stabilization
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Solution Overview
Problem
Conventional nucleic acid sequencing methods face challenges with poor loading uniformity and stability of nucleic acids on solid supports, leading to issues such as nucleic acid detachment and structural instability during analysis.
Innovation Solution
A method involving the use of a poloxamer-containing reagent to treat the solid support, combined with partially double-stranded oligonucleotides to hybridize with nucleic acids before or after loading, enhances the stability and uniformity of nucleic acid attachment.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional loading methods are used to attach nucleic acid to solid support, then the process is simple, but the loading uniformity and stability are poor
Solution Approach 1:
The patent introduces poloxamer as an intermediary substance to facilitate the attachment of nucleic acid nanoballs to the solid support. The poloxamer-containing reagent acts as a mediator that improves the loading uniformity by enabling more consistent and stable attachment of nucleic acid structures to the chip surface, thereby resolving the contradiction between simple process and poor loading uniformity.
Solution Approach 2:
The patent modifies the chemical parameters of the loading environment by introducing poloxamer-containing reagents with specific surfactant properties. This parameter change in the chemical composition of the loading buffer improves the loading uniformity and stability of nucleic acid on the solid support without significantly complicating the overall process.
2Reliability
If conventional loading methods are used, then the process is straightforward, but the stability of nucleic acid after loading is poor
Solution Approach 1:
Poloxamer serves as a stabilizing intermediary between the nucleic acid nanoballs and the solid support surface. This surfactant-containing reagent forms a protective interface that enhances the stability of attached nucleic acid, preventing detachment and degradation during subsequent sequencing operations, while adding only a simple treatment step to the process.
3Measurement precision
If poloxamer-containing reagent is used to improve loading uniformity, then signal-to-noise ratio improves, but the process becomes more complex
Solution Approach 1:
The patent modifies the chemical parameters of the loading environment by incorporating poloxamer-containing reagents. This parameter change in the buffer composition leads to improved loading uniformity, which directly enhances the signal-to-noise ratio in subsequent sequencing measurements. The process complexity increase is minimal, involving only the addition of this specialized reagent to the loading protocol.
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
Improves signal-to-noise ratio and fidelity of nucleic acid analysis by stabilizing nucleic acids on the solid support, reducing signal attenuation and enhancing the integrity of nucleic acid structures during sequencing.
Implementation Method 1
treating the solid support with a hydrophilic poloxamer... significantly improve the loading of nucleic acids (such as DNA nanoballs (DNBs)) on the solid support
Implementation Method 2
contacting the nucleic acid molecule with a partially double-stranded oligonucleotide to hybridize the nucleic acid molecule with the oligonucleotide before or after loading the nucleic acid on the solid support
Data Source
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AI summary
The present invention provides a method for improving the loading of nucleic acid on a solid support by contacting the solid support with a poloxamer-containing reagent. The present invention also provides a method for improving the stability of a nucleic acid on a solid support, comprising contacting a nucleic acid molecule with a partially double-strand oligonucleotide before or after loading the nucleic acid molecule on a solid support, so as to cause the nucleic acid molecule to hybridize with the oligonucleotide. The present invention also provides a combined use of the two methods.