Target Enrichment via Preliminary Fragmentation and Probe Hybridization
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Solution Overview
Problem
Current methods for analyzing genes require the creation of a DNA library, which is time-consuming, results in sample loss, and generates fragments with undefined locations, making it inefficient for enriching target sequences.
Innovation Solution
The method involves randomly fragmenting nucleic acids, adding target probes and enzymes to isolate specific sequences, and creating a target-enriched library by removing non-target nucleic acids, allowing for unbiased coverage of target regions for further studies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If a DNA library is created prior to enrichment with target probes, then target sequences can be enriched, but the process is time-consuming and results in sample loss
Solution Approach 1:
The method performs preliminary fragmentation of the nucleic acid sample before library creation and enrichment steps. By fragmenting the sample first and then using target probes to capture and enrich specific sequences directly from the fragmented pool, the workflow eliminates the time-consuming step of creating a complete library before enrichment, while still achieving reliable target sequence enrichment
Solution Approach 2:
The invention extracts only the necessary components for target enrichment by using target probes to selectively capture and isolate specific nucleic acid sequences from the fragmented sample pool. This extraction approach avoids the need to process and store complete libraries, reducing time and sample loss while maintaining enrichment effectiveness
2Reliability
If a DNA library is created prior to enrichment, then target sequences can be captured, but sample loss occurs and amplification is required
Solution Approach 1:
The method extracts and isolates only the target sequences of interest from the fragmented nucleic acid sample using complementary target probes. By directly capturing specific sequences from the fragmented pool without creating a complete library first, the process minimizes sample loss and eliminates the need for extensive amplification steps
Solution Approach 2:
Target probes serve as intermediaries that mediate the selective capture and enrichment of specific nucleic acid sequences from the fragmented sample. These probes hybridize to complementary target sequences, enabling specific isolation and concentration of target regions without requiring library construction and reducing overall sample consumption
3Reliability
If a DNA library is created with target probes, then enrichment can occur, but target regions are at undefined locations within fragments
Solution Approach 1:
The method performs preliminary fragmentation of the nucleic acid sample into smaller pieces before applying target probes for enrichment. This preliminary fragmentation, combined with subsequent target-specific capture, ensures that target sequences are present in defined, recoverable fragments rather than at undefined locations within large library constructs, facilitating precise isolation and downstream analysis
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 efficiently enriches target DNA/RNA sequences, reducing sample loss and time, while ensuring target sequences are isolated and amplified accurately, suitable for gene expression studies and disease-specific transcript identification.
Implementation Method 1
The nucleic acid sample is randomly fragmented into a pool of DNA fragments using physical or enzymatic methods
Implementation Method 2
The nucleic acid sample is randomly fragmented into a pool of DNA fragments using physical or enzymatic methods
Implementation Method 3
The fragmented nucleic acid pool is hybridized with target probes or baits to capture and enrich the target sequences
Data Source
AI summary
The present disclosure provides a method and a kit for selecting and enriching target sequences specific for one or more genomic regions of interest or a subset of a transcriptome using a target-capturing library of probes. The target-enriched library is generated from a random pool of deoxyribonucleic acid (DNA) fragments. The present disclosure provides an efficient and cost-effective method of target selection for targeted genome sequencing, targeted nucleic acid library creation and gene expression studies.