Single-Assay DNA and RNA Library Generation on PolyT Flowcells
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Solution Overview
Problem
Traditional methods for analyzing multiple analytes in a biological sample require separate assays, increasing time, cost, and resource consumption, and do not efficiently utilize the sample's comprehensive information.
Innovation Solution
A system and method for simultaneously analyzing DNA and RNA in a single assay using a flowcell device with polyT capture probes that hybridize to RNA and DNA, allowing for the generation of cDNA and gDNA libraries in a single compartment, and utilizing transposase-based tagmentation for indexing and capturing analytes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If separate assays are used to analyze different analytes (DNA, RNA, proteins), then each analyte can be analyzed with dedicated reagents and protocols, but the overall analysis time, cost, and resource consumption increase significantly
Solution Approach 1:
The patent combines multiple analyte analysis (DNA, RNA, proteins) into a single multiplexed assay performed in a single well. Different analytes are simultaneously detected using a shared reagent system with analyte-specific capture sequences, eliminating the need for separate assays and reducing total analysis time while maintaining detection accuracy for each analyte type
Solution Approach 2:
The invention creates a universal assay platform that can detect multiple analyte types using a common reagent formulation. The reagent system includes capture sequences that can specifically bind to different analytes (DNA, RNA, proteins) within the same reaction mixture, allowing one reagent system to perform multiple detection functions simultaneously
2Adaptability or versatility
If multiple separate assays are performed for comprehensive analyte analysis, then complete coverage of all analytes is achieved, but the complexity of the overall process increases
Solution Approach 1:
The patent implements a universal reagent system that maintains versatility for detecting multiple analyte types (DNA, RNA, proteins) while reducing assay complexity. The same reagent formulation with differentiated capture sequences handles all analyte types in a single well, eliminating the need for multiple separate assay setups and reducing procedural complexity
3Reliability
If separate assays are used for each analyte, then specific reagents can be optimized for each target, but resource consumption and cost increase
Solution Approach 1:
The patent merges reagent usage across multiple analyte detections into a single shared reagent system. The reagents are designed with analyte-specific capture sequences that enable selective binding to different targets (DNA, RNA, proteins) within the same reaction well, reducing reagent consumption while maintaining detection reliability through specific analyte-reagent interactions
Solution Approach 2:
The invention creates universal reagents that can detect multiple analyte types with equal reliability. The reagent system includes capture sequences specifically designed to bind different analytes, allowing one reagent formulation to replace multiple specialized reagents while maintaining the same level of detection reliability for each analyte type
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 reduces assay complexity and time, enhances analysis efficiency, and enables the simultaneous generation of multiple nucleic acid libraries, improving the cost-effectiveness and accuracy of analyzing multiple analytes.
Implementation Method 1
a polyT capture probe configured to hybridize to a 3' polyA tail of RNA and a 3' polyA tail that has been tagmented to DNA
Implementation Method 2
utilizing transposase-based tagmentation for indexing and capturing analytes
Data Source
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AI summary
Embodiments of systems, methods, and compositions provided herein relate to methods of simultaneously analyzing multiple analytes in a single sample using a single assay. Some embodiments relate to simultaneous analysis of DNA and RNA in a single sample, for example, to the simultaneous generation of DNA and RNA libraries.