Pre-capture Multiplexing: An Efficient Strategy for NGS
Next-generation sequencing (NGS) technology enables the cost-effective generation of vast amounts of sequencing data, making it widely applicable across various fields. Given the high data output produced in a single run, index sequences are used to distinguish data from multiple samples analyzed simultaneously.
By preparing indexed libraries for each sample and pooling them before the target enrichment reaction, NGS experiments can be conducted more efficiently and cost-effectively.
Compared to performing capture for individual samples, pre-capture multiplexing offers several advantages:
- Cost Reduction: Lowers reagent consumption, making the process more economical.
- Reduced Experimental Time: Pooling multiple samples into a single reaction reduces overall hands-on time.
- Consistent Data Coverage: Since pooled samples undergo the same hybridization capture reaction in a single tube, variations caused by handling are minimized, ensuring uniform coverage.
- Efficient Use of Low-Input Samples: When the library quantities are insufficient for hybridization capture, pooling multiple samples enables successful data acquisition even with limited sample input.
However, improper mixing ratios may result in insufficient sequencing data for certain samples. Additionally, the presence of low-quality libraries can negatively affect the results of all pooled samples.
Single-plex, 12-plex Pre-capture Multiplexing Target Enrichment Results
With years of experience in Target Enrichment panel development, Celemics has optimized panels to maintain stable Target Enrichment efficiency for up to 12-plex multiplexing. The graph below presents internal validation data, comparing the results of Single-plex and 12-plex pre-capture multiplexing, confirming that library data quality remains consistent with minimal variation.
Celemics provides a range of hybridization-based target capture solutions, including Single-plex, 4-plex, 8-plex, and 12-plex, to accommodate diverse research needs.
