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  5. Next Gen Sequencing Kits
  6. NGS library quantification kit

NGS library quantification kit

o 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.

Brand: Takara

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Product CodeProduct Product Description Price (Excl gst) Qty Add
CLT638325 DNA Standards for Library Quantification, 50 Rxns DNA Standards for Library Quantification, 50 Rxns Login for price  
CLT638324 Library Quantification Kit, 500 Rxns Library Quantification Kit, 500 Rxns Login for price  

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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

Applications

  • Illumina NGS library quantification by qPCR

The DNA Standards for Library Quantification includes four DNA standards (0.01–10 pM) that are used for qPCR-based library quantification prior to Illumina next-generation sequencing. These DNA standards contain specific concentrations of a DNA fragment (538 bp) and are designed specifically for use with the Library Quantification Kit (Cat No. 638324). When used as templates for qPCR with the Library Quantification Kit, the standards will generate 447-bp DNA fragments that can be used to produce a standard curve for determining the concentration of DNA in an Illumina library. The standards are not suitable for use with other qPCR techniques, as they may lead to quantification errors or poor results.

The Library Quantification Kit is used for rapid library quantification prior to Illumina next-generation sequencing. The kit contains all of the components needed to determine the amount of library DNA (genomic, cDNA, etc.) in a sample using qPCR. Quantification is accomplished by comparison of the threshold cycle (Ct) of the diluted sample with a standard curve derived from four calibrated DNA standards.