KAPA Library Quantification Kits

KAPA Library Quantification Kits

Next-generation DNA Sequencing meets Next-generation qPCR

Current standard protocols for commercial next generation sequencing platforms ­employ laborious, costly, and often unreliable methods for quantifying DNA libraries. 

Accurate quantification of PCR-competent sequencing templates is crucial for reliable clonal amplification via either emulsion PCR (emPCR) or bridge PCR (bPCR) – ­underestimation usually results in non-clonality, while overestimation can lead to inefficiency via poor yields of clonally amplified templates. 

Standard methods for quantifying NGS ­libraries have a number of important disadvantages. Electrophoresis and spectrophotometry measure total nucleic acid concentrations, whereas optimal cluster density or template-to-bead ratio depend on the appropriate concentration of PCR-amplifiable DNA molecules. These methods also have low sensitivity, consuming nanograms of precious samples, and are not suitable for high-throughput workflows. Quantitative PCR (qPCR) is inherently ­well-suited for next-generation sequencing library quantification: 

  • qPCR specifically quantifies only PCR-competent DNA molecules 
  • is highly sensitive allowing accurate quantification of low concentration libraries

 

Your benefit

 

  • Reliable and sensitive quantification of all sequencing-competent library molecules 
  • Accurate and reproducible quantitation across a wide range of library types, concentrations, fragment length distributions and GC content 
  • More efficient, equimolar pooling for ­multiplexed sequencing 
  • Flexibility to support manual and automated, high-throughput pipelines; as well as PCR-free workflows 

 

Product characteristics

 

qPCR library quantification results in streamlined workflows 
KAPA Library Quantification Kits eliminate the need for time-consuming and expensive titrations and provide a conducive ­format for streamlining high-throughput workflows.

Reliable quantification results in consistent cluster density 

Efficient amplification of a wide range of templates during qPCR 
Traditional qPCR reagents are optimized for short amplification targets; longer targets, unbalanced GC-content, and problematic secondary structures may result in low ­amplification efficiency and unreliable quantification of some library molecules. 
To address the demands of quantifying complex DNA libraries, Kapa Biosystems has engineered a DNA polymerase specifically for SYBR® Green-based qPCR, enabling efficient amplification of targets that present a challenge to wild-type enzymes. KAPA Library Quantification Kits contain this engineered polymerase to ensure ­robust amplification of longer fragments, across a broad range of GC-content, required for accurate library quantification. 

Reliable DNA quantification standards with minimal variability from lot-to-lot

 


qPCR Library Quantification results in streamlined workflows

 

 

 

For life science research only. 
Not for use in diagnostic procedures.

Data on file.