Next-Generation DNA Sequencing

From Sample Enrichment to Genome Finishing: Kapa Biosystems offers a range of products to improve your NGS workflow.

NEW! KAPA Library Quantification Kits

Eliminate the need for expensive and time-consuming titrations – use qPCR as a fast and reliable method for quantifying libraries for the Roche 454 FLX/Titanium and Illumina Genome Analyzer next-generation sequencing platforms. 
 
Accurate quantification of PCR-competent sequencing templates is crucial for reliable clonal amplification via either emulsion PCR (emPCR) or bridge PCR (bPCR) - underestimation results in non-clonality, while overestimation leads to inefficiency via poor yields of clonally amplified templates.
 
KAPA Library Quantification Kits combine a quality-controlled set of DNA quantification standards with the unmatched performance of KAPA SYBR® FAST qPCR reagents to provide a rapid, sensitive, and reliable method for quantifying amplifiable molecules in next-generation sequencing DNA libraries.
 
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.
 
AGBT 2010 Poster: The Implementation of KAPA Library Quantification Kits at the Broad Institute Leads to Streamlined Workflows and Reduced Cluster Variability.

Targeted Sample Enrichment

Achieve efficient and consistent coverage of your region of interest with KAPA Long Range HotStart DNA Polymerase. High yield amplification from complex targets up to 20 kb with minimal optimization. 
 
Pooled Amplicon Libraries for Resequencing
The ultra-high fidelity and high yields of KAPAHiFi HotStart DNA Polymerase is critical for minimizing PCR errors and increasing coverage when pooling amplicon libraries for resequencing projects using next-generation DNA sequencing.   

Genome Closure and Finishing

The relatively short read lengths of NGS platforms make whole genome assembly more problematic and result in more sequence gaps than traditional Sanger sequencing methods. Without subclones, next-generation whole genome sequencing requires PCR for finishing and sequence gap closure. KAPAHiFi HotStart is recommended when finishing PCR products are sequenced using NGS. Ultra-high fidelity is important since all NGS templates are clonally amplified and are therefore more sensitive to errors introduced by PCR.
 
If the sequence context around the gaps is difficult to PCR due to high GC content or secondary structure, KAPA2G Robust HotStart is recommended. The higher processivity of the engineered KAPA2G Robust DNA Polymerase makes this the enzyme of choice for amplifying difficult templates.