Barcode Oligonucleotides for Sample Multiplexing in Genomic Sequencing
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Solution Overview
Problem
Current methods for analyzing genomic differences across multiple samples are costly and inefficient, as they require separate processing of each sample, limiting throughput and increasing costs.
Innovation Solution
The method involves attaching unique identifiers to reactant molecules from different samples, allowing them to be pooled and then separated post-analysis, enabling correlation of data back to their original sample through barcode oligonucleotides, facilitating sample multiplexing and sequencing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If separate processing of each sample is performed, then sample identification accuracy is maintained, but throughput is limited and costs increase
Solution Approach 1:
Multiple samples are pooled together into a single reaction mixture, allowing simultaneous processing of multiple samples in one sequencing run. This merging approach increases throughput while maintaining sample identification through unique barcodes attached to each sample's nucleic acid molecules.
Solution Approach 2:
Unique barcode oligonucleotides serve as intermediary markers that are attached to nucleic acid molecules from different samples. These barcodes enable tracking and identification of molecules throughout the pooled processing workflow, allowing data to be correctly attributed to original samples after multiplexed analysis.
2Loss of substance
If separate processing of each sample is performed, then sample-specific analysis is ensured, but reagent costs and preparation time increase
Solution Approach 1:
Multiple samples are combined into a single pooled reaction, allowing shared use of reagents such as enzymes, buffers, and sequencing chemicals across all samples. This dramatically reduces per-sample reagent consumption and eliminates redundant preparation steps for each individual sample.
Solution Approach 2:
Unique barcodes are attached to nucleic acid molecules from different samples in a preliminary step before pooling. This advance labeling enables subsequent multiplexed processing without loss of sample identification capability, reducing overall preparation time by enabling parallel processing of multiple samples simultaneously.
3Productivity
If samples are pooled together, then throughput and cost efficiency improve, but sample identification becomes more challenging
Solution Approach 1:
Unique barcode oligonucleotides act as information carriers that are attached to nucleic acid molecules from specific samples. These barcodes preserve sample origin information throughout the pooling and processing workflow, enabling accurate reconstruction of which molecules originated from which samples after multiplexed analysis.
Solution Approach 2:
Each sample pool is赋予 (endowed with) a unique barcode sequence that distinguishes it from other samples. This localized quality marker on specific molecules allows the system to maintain individual sample identity within the heterogeneous pooled mixture, preventing information loss despite physical mixing of samples.
Data Source
AI summary
The invention generally relates to methods for sample multiplexing. In certain embodiments, methods of the invention obtaining a plurality of different reactant molecules, attaching a unique identifier to the reactant molecules, and forming a droplet including the reactant molecules.