NGS library quantification kit
To obtain high-quality Illumina next-generation sequencing (NGS) data, loading the flow cell with an appropriate amount of library is essential. Loading an insufficient amount of library will result in low cluster density and reduced sequencing yield; an overabundance of library may increase cluster density and result in poor-quality data. Standard methods of NGS library quantification, such as those based on electrophoresis or spectrophotometry, have low sensitivity, are non-specific for adapter-bound DNA, and typically require a large amount of library sample for analysis. The Library Quantification Kit provides a highly sensitive, qPCR-based quantification method that specifically targets adapter sequences used for Illumina's NGS reactions. This kit allows specific measurement of the sequencing library concentration even in samples that contain DNA molecules not bound to adapters. This kit can also be used to confirm adapter ligation during library preparation.
To obtain high-quality Illumina next-generation sequencing (NGS) data, loading the flow cell with an appropriate amount of library is essential. Loading an insufficient amount of library will result in low cluster density and reduced sequencing yield; an overabundance of library may increase cluster density and result in poor-quality data. Standard methods of NGS library quantification, such as those based on electrophoresis or spectrophotometry, have low sensitivity, are non-specific for adapter-bound DNA, and typically require a large amount of library sample for analysis. The Library Quantification Kit provides a highly sensitive, qPCR-based quantification method that specifically targets adapter sequences used for Illumina's NGS reactions. This kit allows specific measurement of the sequencing library concentration even in samples that contain DNA molecules not bound to adapters. This kit can also be used to confirm adapter ligation during library preparation.
Library quantification with this kit is accomplished by qPCR using TB Green for detection. Amplification products are detected by intercalation of TB Green I in double-stranded DNA. Fluorescence detection in real time allows quantification of amplification products. This kit includes DNA standards of known concentration to allow absolute quantification of library DNA.
Overview
- Highly sensitive quantification of Illumina sequencing libraries—the qPCR-based assay can measure libraries at low concentrations
- Allows specific quantification of only adapter-bound DNA molecules
- Reduces variability in cluster density to improve sequencing results—use prior to sequencing to ensure optimal loading concentration
- Complete kit—includes qPCR primers and reagents, and DNA standards for absolute quantification using a standard curve
Interested in more data and FAQs about this product? Visit the NGS Learning Center.
More Information
Applications
- Illumina NGS library quantification by qPCR
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
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