Degradable Barcode Beads for Genomic Sample Processing
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Solution Overview
Problem
Current genomic sequencing methods face challenges in efficiently processing and identifying nucleic acid samples, particularly in diagnostics, biotechnology, and forensic biology, due to limitations in sample processing and identification techniques.
Innovation Solution
The development of methods and compositions for generating beads with covalently attached polynucleotides, which involve barcoding sample materials by releasing nucleic acid barcode molecules from a first partition into a second partition, allowing for the attachment of these barcodes to sample components, enabling efficient sample identification and sequencing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional sample processing methods are used, then sample identification can be performed, but processing efficiency and precision are insufficient
Solution Approach 1:
The sample processing system is segmented into multiple partitions, each containing barcode molecules that can be selectively released and attached to specific sample components. This segmentation enables parallel processing of multiple samples simultaneously while maintaining precise identification through unique barcode sequences associated with each partition.
Solution Approach 2:
Barcode molecules serve as intermediary elements that bridge the sample components and the identification system. These barcodes are released from partitions and attached to sample components, enabling efficient barcoding and subsequent precise identification through sequencing while improving overall processing throughput.
2Productivity
If barcode molecules are released from partitions, then sample barcoding efficiency improves, but control over the release process becomes more complex
Solution Approach 1:
The release of barcode molecules from partitions is controlled by changing physical or chemical parameters such as temperature, pH, or ionic strength. By adjusting these parameters, the barcode molecules can be selectively released from partitions under controlled conditions, improving barcoding efficiency while maintaining manageable process complexity through standardized parameter changes.
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 precise and efficient barcoding of sample materials, facilitating advanced nucleic acid analysis and sequencing by allowing for the attachment of barcode sequences to sample components, thereby improving sample identification and processing efficiency in genomic sequencing applications.
Implementation Method 1
the barcode molecules may be released from the first partition by degrading the first partition
Implementation Method 2
cleaving a chemical linkage between the barcode molecules and the bead
Implementation Method 3
exposing the bead to a reducing agent (e.g., dithiothreitol (DTT) or tris(2-carboxyethyl)phosphine (TCEP))
Implementation Method 4
at least one of crosslinking of the bead and a linkage between the bead and the barcode molecules may comprise a disulfide linkage
Implementation Method 5
generating beads with covalently attached polynucleotides
Data Source
AI summary
This disclosure provides methods and compositions for sample processing, particularly for sequencing applications. Included within this disclosure are bead compositions, such as diverse libraries of beads attached to large numbers of oligonucleotides containing barcodes. Often, the beads provides herein are degradable. For example, they may contain disulfide bonds that are susceptible to reducing agents. The methods provided herein include methods of making libraries of barcoded beads as well as methods of combining the beads with a sample, such as by using a microfluidic device.