SeqCap EZ HyperCap Workflow

New SeqCap EZ HyperCap User's Guide v1.0  

Performance. Speed. Automatability.  

For use across all SeqCap EZ Target Enrichment products

The SeqCap EZ HyperCap Workflow provides an integrated workflow solution that combines high-efficiency library preparation kits from KAPA with performance-optimized SeqCap capture protocol. It is an application-specific approach for efficient, streamlined and automatable targeted sequencing.

KAPA Hyper Prep and KAPA HyperPlus kits are novel one-tube chemistry with optimally formulated and evolved enzymes that enable higher yields of adapter-ligated library and lower amplification bias resulting in higher library diversity, lower duplication rates and more uniform coverage. 

SeqCap EZ Target Enrichment System is an innovative single-tube solution-based hybridization capture technology that enables enrichment of any genomic region of interest. Built upon an optimized design algorithm and manufactured with high probe density, the SeqCap EZ target enrichment products demonstrate uniform and higher depth of coverage, enabling discovery and detection of variants. 

Benefits of the HyperCap protocol include:

Speed with streamlined workflow1

  • Hybridization-based capture with enzymatic or mechanical-shearing protocol
  • Greatly reduced DNA fragmentation and library construction time in comparison to commonly used library preparation protocols  

Versatile performance for maximum flexibility2

  • Further improved enrichment metrics than KAPA Library Preparation Kits (Figure 1a & 1b)
  • Streamlined protocol demonstrating comparable coverage metrics as standard solution (Figure 2)

Easily automatable workflow with broad platform compatibility 3

  • Reduced instrumentation requirement enabling easy automation (Table 1)
  • Workflow allows multiplexing, thus optimizing robot utilization
  1. Data on file
  2. SeqCap EZ HyperCap Workflow User’s Guide v1.0
  3. See KAPA automation site

Performance data using SeqCap EZ HyperCap Workflow v1.0

Performance metrics using SeqCap EZ MedExome Target Enrichment Kit and SeqCap EZ Comprehensive Cancer Panel Design using SeqCap EZ HyperCap Workflow v1.0 was compared to the existing SeqCap EZ SR User's Guide v5.1.

Note: The SeqCap EZ SR User's Guide v5.1 utilizes the standard KAPA Library Prepration Kits, not the KAPA Hyper Prep or KAPA HyperPlus kits. 

MedExome-Uniformity-DupRate

Figure 1a

Cancer-Panel-Uniformity-DupRate

Figure 1b

Figure 1a & 1b: Total PCR duplicate rate and Fold 80 base penalty. Uniformity is measured as Fold 80 base penalty for the SeqCap EZ MedExome and the SeqCap EZ Comprehensive Cancer Panel Design using the SeqCap EZ HyperCap Workflow User’s Guide v1.0.  

Cancer-Panel-Enrichment-Metrics

Figure 2

Figure 2: Enrichment metrics for the Comprehensive Cancer Panel Design using SeqCap EZ HyperCap Workflow User’s Guide v1.0. The new SeqCap EZ HyperCap protocol using either KAPA Hyper Prep or KAPA HyperPlus kits showed comparable performance against the standard KAPA Library Preparation Kit recommended by SeqCap EZ User’s Guide v5.1. Coverage (at >20X) improved with the HyperCap protocol.

Experiment Details: 100 ng of NA12891 Coriell DNA was evaluated with SeqCap EZ MedExome and Comprehensive Cancer Panel using one of the three KAPA library preparation Kits following appropriate SeqCap EZ protocol. For the HyperCap protocol, either the KAPA Hyper Prep kit or the KAPA HyperPlus Kit was included for library construction. Following library preparation, the libraries were processed for capture. The enriched libraries were sequenced. . The sequencing reads were randomly sampled to 60 million reads (SeqCap EZ MedExome Kit) or 4 million reads (Comprehensive Cancer panel) for each sample and analyzed using informatics pipeline as discussed in Tech Note
"How to evaluate SeqCap EZ enrichment data". 

 

SeqCap EZ HyperCap Workflow enables easy automation

Automation-Table

Table 1

Table1:  Automation platforms available for use with SeqCapEZ enrichment products.
Note: The availability of “Standard Solutions” is constantly changing. Contact your instrument providers or see KAPA's Automation webpage to obtain the most up-to-date list of available applications. 

Learn more about SeqCap EZ HyperCap Workflow

SeqCap EZ HyperCap Workflow User’s Guide v1.0

For additional information, please contact your Roche sales representative. 

For Research Use Only. Not for use in diagnostic procedures.