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KAPA HyperPrep Kits offer a streamlined library preparation protocol that combines several enzymatic steps and eliminates bead cleanups to significantly reduce library preparation time and improve consistency. The novel, single-tube chemistry offers further improvements to library construction efficiency, particularly for challenging samples such as FFPE and cell-free DNA. KAPA HyperPrep Kits offer complete library preparation solution with KAPA Adapters and KAPA Pure Beads.
Features and Benefits of KAPA HyperPrep Kits
Robust and reproducible library construction in 2-3 hours with high conversion rates and library complexity, particularly for FFPE
PCR-free workflows from lower sample inputs and improved sequence coverage for workflows with PCR
Compatibility with a wide range of sample types and inputs, and flexibility with respect to fragment size, adapter design and library amplification
Higher library yields translating to greater molecular complexity
Higher conversion of input DNA to adapter-ligated library
Fewer cycles of amplification with KAPA HiFi DNA Polymerase, resulting in lower duplication rates and improved coverage
View comparison data for higher conversion of input DNA to adapter-ligated library by KAPA HyperPrep Kit compared to Illumina TruSeq Nano DNA Kits or NEBNext® Ultra™ II FS Kit.
Figure 1. KAPA HyperPrep Kits enable higher library construction efficiency. Conversion rates for libraries prepared for target capture from different amounts of Covaris-sheared human genomic DNA, using the KAPA HyperPrep or competitor library construction kits. Libraries were prepared according to manufacturer’s instructions. Input DNA was quantified by Qubit, whereas the qPCR-based KAPA Library Quantification Kit was used to determine KAPA HyperPrep and TruSeq Nano library yields after adapter ligation. Conversion rates for NEBNext libraries cannot be measured directly with the KAPA Library Quantification Kit and were derived from post-amplification yields.
Minimal amplification bias
Kits with or without an amplification module
Lower PCR bias resulting in more uniform coverage in PCR workflows
See performance data showing high coverage uniformity using KAPA HyperPrep Kits and minimal amplification bias with KAPA HiFi polymerase.
Figure 2. KAPA HiFi introduces minimal amplification bias and offers high coverage uniformity. GC bias plots for libraries prepared for whole-genome shotgun sequencing of bacteria with extreme genomic GC content (Clostridium, 29% GC and Bordetella, 68% GC). Libraries were prepared with the KAPA HyperPrep Kit from 100 ng or 1 ng DNA, Covaris-sheared to an average size of ~200 bp; and sequenced without amplification, or after amplification with KAPA HiFi. Minimal amplification bias was introduced by KAPA HiFi, despite a relatively high number of PCR cycles (12 cycles for Clostridium and 13 cycles for Bordetella). Paired-end sequencing (2 x 300 bp) was performed on an Illumina MiSeq, and data analyzed using Picard.
Figure 3. KAPA HiFi introduces minimal amplification bias and offers high coverage uniformity. GC bias plots for libraries prepared for whole-genome shotgun sequencing of bacteria with extreme genomic GC content (Clostridium, 29% GC and Bordetella, 68% GC). Libraries were prepared with the KAPA HyperPrep Kit from 100 ng or 1 ng DNA, Covaris-sheared to an average size of ~200 bp; and sequenced without amplification, or after amplification with KAPA HiFi. Minimal amplification bias was introduced by KAPA HiFi, despite a relatively high number of PCR cycles (12 cycles for Clostridium and 13 cycles for Bordetella). Paired-end sequencing (2 x 300 bp) was performed on an Illumina MiSeq, and data analyzed using Picard.
Higher library yield, higher library complexity and fewer PCR cycles
Lower duplication rates and higher coverage
View data showing higher library diversity, lower duplication rates and improved coverage and using KAPA HyperPrep Kit in comparison with TruSeq Nano DNA Kit.
Figure 4. KAPA HyperPrep Kits enable improved coverage depth for FFPE samples. Key sequencing metrics for libraries prepared from 100 ng of FFPE DNA for target capture, using either the KAPA HyperPrep Kit or TruSeq Nano DNA Sample Prep Kit (Illumina). Captures were performed with the SeqCap EZ Comprehensive Cancer Design (4 Mb) according to the manufacturer’s instructions, with the exception that the number of pre-capture amplification cycles for each library type was optimized based on post-ligation yields (10 cycles for KAPA HyperPrep vs. 14 cycles for TruSeq Nano DNA libraries). All libraries were amplified for 13 cycles after capture. More efficient library construction and less PCR bias with the KAPA HyperPrep Kits resulted in higher library complexity prior to capture, lower duplication rates, and improved coverage. Sequencing (2 x 75 bp) was performed on an Illumina HiSeq 2500. Sequencing reads were down-sampled to ~14 million per library prior to analysis with Picard.
Figure 5. KAPA HyperPrep Kits enable improved coverage depth for FFPE samples. Key sequencing metrics for libraries prepared from 100 ng of FFPE DNA for target capture, using either the KAPA HyperPrep Kit or TruSeq Nano DNA Sample Prep Kit (Illumina). Captures were performed with the SeqCap EZ Comprehensive Cancer Design (4 Mb) according to the manufacturer’s instructions, with the exception that the number of pre-capture amplification cycles for each library type was optimized based on post-ligation yields (10 cycles for KAPA HyperPrep vs. 14 cycles for TruSeq Nano DNA libraries). All libraries were amplified for 13 cycles after capture. More efficient library construction and less PCR bias with the KAPA HyperPrep Kits resulted in higher library complexity prior to capture, lower duplication rates, and improved coverage. Sequencing (2 x 75 bp) was performed on an Illumina HiSeq 2500. Sequencing reads were down-sampled to ~14 million per library prior to analysis with Picard.
Ability to optimize adapter:insert ratios for challenging sample types, such as cell-free DNA
Higher library density compared to competitor kit*
Comparable duplication rates to vendor kits specific for low-input applications
See comparative data for improved library diversity using KAPA HyperPrep Kit compared to competitor kit. View data showing comparable duplication rates by KAPA HyperPrep Kit as those by a competitor kit designed specifically for low-input applications.
Figure 6. KAPA HyperPrep Kits enable improved sequencing metrics for libraries prepared from cell-free DNA. Library diversity and duplication rates for libraries prepared from 2 ng of cell-free DNA, using the KAPA HyperPrep Kit or a competitor low-input library construction kit (blue). Libraries were prepared according to manufacturer’s instructions. KAPA HyperPrep libraries were prepared with a range of adapter:insert molar ratios and the adaper:insert ratio of the competitor kit was unknown. Sequencing (2 x 150 bp) was performed on an Illumina MiSeq and data analyzed using Picard.
Figure 7. KAPA HyperPrep Kits enable improved sequencing metrics for libraries prepared from cell-free DNA. Library diversity and duplication rates for libraries prepared from 2 ng of cell-free DNA, using the KAPA HyperPrep Kit (green) or a competitor low-input library construction kit (red). Libraries were prepared according to manufacturer’s instructions. KAPA HyperPrep libraries were prepared with a range of adapter:insert molar ratios and the adaper:insert ratio of the competitor kit was unknown. Sequencing (2 x 150 bp) was performed on an Illumina MiSeq and data analyzed using Picard.
The All of Us Research Project is a groundbreaking initiative led by the National Institutes of Health (NIH) in the United States. The aim of this project is to accelerate research and improve health outcomes for everyone by gathering data from at least one million people; approximately 77% of these people are from previously under-represented groups. To prepare these important samples for sequencing, NIH chose KAPA HyperPrepLibrary Preparation Kits; the robust chemistry of these kits enabled the reliable,consistent preparation of samples across multiple research sites.
Find NGS library primer mixes and amplification kits with enzymes specially engineered for low-amplification bias and uniform coverage of difficult regions.
Using a qPCR-based solution, KAPA Library Quantification Kits provide accurate and reliable quantification of libraries prepared for sequencing on Illumina and IonTorrent platforms, across a wide range of library types, concentrations, fragment distributions and GC content. Kits include KAPA SYBR FAST qPCR Master Mix, a platform-specific library quantification primer premix, and a pre-diluted set of DNA standards.
KAPA Stranded RNA-seq Kits are our first generation RNA library preparation kits that combine a “with-bead” protocol with the robustness of KAPA HiFi DNA Polymerase to yield high-quality RNA-seq libraries.
Learn more about our streamlined and automation-friendly workflow solution. It integrates KAPA Library Preparation Kits and KAPA HyperPlex Adapters with KAPA Target Enrichment Probes, Roche Universal Enhancing Oligos and our KAPA HyperPure Beads.
KAPA Single-Indexed Adapter Set A contains indices 2, 4, 5, 6, 7, 12, 13, 14, 15, 16, 18 and 19, whereas Set B contains indices 1, 3, 8, 9, 10, 11, 20, 21, 22, 23, 25, 27. All KAPA Single- and Dual-Indexed Adapter Kits contain KAPA Adapter Dilution Buffer. KAPA Dual-Indexed Adapter Kits also contain three additional sealing films to support multiple use.
KAPA Single-Indexed Adapter Set A contains indices 2, 4, 5, 6, 7, 12, 13, 14, 15, 16, 18 and 19, whereas Set B contains indices 1, 3, 8, 9, 10, 11, 20, 21, 22, 23, 25, 27. All KAPA Single- and Dual-Indexed Adapter Kits contain KAPA Adapter Dilution Buffer. KAPA Dual-Indexed Adapter Kits also contain three additional sealing films to support multiple use.
KAPA Code
Roche Cat. No
Description
Kit Size
How to buy
KK8000
07983271001
KAPA Pure Beads (5 mL)
5 mL
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KK8001
07983280001
KAPA Pure Beads (30 mL)
30 mL
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KK8002
07983298001
KAPA Pure Beads (60 mL)
60 mL
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KK8721
08278539001
KAPA Adapter Dilution Buffer (25 mL)
25 mL
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KK8727
08861919702
KAPA Unique Dual-Indexed Adapter Kit, (15 µM)
96 adapters x 20 µl each
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Additional pack sizes are available for KAPA HyperPure Beads. Please speak to your local sales representative.
