Product List
Library Quantification
KAPA Library Quantification Kits contain all the reagents needed for the accurate, reliable and reproducible qPCR-based quantification of next-generation sequencing (NGS) libraries prepared for sequencing on Illumina and IonTorrent platforms. Kits include KAPA SYBR FAST qPCR Master Mix (formulated with different passive reference dyes for different qPCR instruments), a platform-specific library quantification primer premix, and a pre-diluted set of DNA standards.
Six pre-diluted DNA Standards and appropriately diluted NGS libraries are amplified using platform-specific qPCR primers that target adapter sequences. The average Cq value for each DNA Standard is plotted against its known concentration to generate a standard curve. The standard curve is used to convert the average Cq values for diluted libraries to concentration, from which the working concentration of each library is calculated.
Benefits
- 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.
- Accurate, equimolar pooling for multiplexed sequencing.
- Flexibility to support manual and automated high-throughput pipelines as well as PCR-free workflows.
- Quantification of only/all sequencing-competent library fragments.
- Use of single DNA standard for all library types.
- Pre-diluted standards with very high lot-to-lot consistency.
Development and validation of a clinical cancer genomic profiling test based on massively parallel DNA sequencing.
DNA Library Preparation
Preparation of a DNA library for next generation sequencing (NGS) platforms entails several steps that have a direct influence on the quality of the sequencing library, and ultimately on the reliability of sequencing results. Roche Sequencing Solutions offers best-in-class* DNA library preparation kits containing high-quality enzymes and reaction buffers developed through our Directed Evolution Technology. The KAPA Library Preparation Kits include modules required for end repair, A-tailing, ligation, and amplification (for Illumina platforms) and offer complete library preparation solutions with KAPA Adapters and KAPA Pure Beads.
KAPA HyperPlus Kits, the most advanced library preparation option from Roche, provide a streamlined workflow that includes fully automatable fragmentation and library preparation in a single tube. Building on industry-leading library construction efficiencies, this integrated solution combines enzymatic fragmentation, similar in quality to mechanical shearing, with the speed and convenience of tagmentation-based workflows. KAPA HyperPlus Kits offer complete library preparation solution with KAPA Adapters and KAPA Pure Beads. The kits are compatible with the Illumina sequencing platform and provide qualified automation methods.
Benefits
- Fully automatable DNA library construction in 1.5–3 hours with low-bias nonmechanical fragmentation.
- Industry-leading conversion rates and library complexity,* particularly for FFPE DNA.
- PCR-free workflows from lower inputs and improved sequence coverage in 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.
KAPA HyperPlus Guide to Success
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.
Benefits
- 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.
- Qualified automation methods.
- Validated with the HyperCap Workflow v2.0.
Clinical utility of circulating tumor DNA for molecular assessment in pancreatic cancer
KAPA HTP/LTP Library Preparation Kits contain optimally formulated enzymes selected through our directed evolution technology and implement a highly optimized "with bead" strategy that enable higher library construction efficiency and facilitate automation. Kits offer higher success rates than conventional library construction kits — particularly from lower inputs and challenging samples such as FFPE and ChIP DNA — in both high- and low-throughput sample preparation pipelines. KAPA HTP/LTP Library Preparation Kits provide a complete library preparation solution with KAPA Adapters and KAPA Pure Beads.
Benefit
- Automation-friendly, highly optimized "with-bead" protocol facilitating minimal sample loss with no physical transfer of library material between enzymatic reactions.
- Improved coverage uniformity as a result of low-bias library amplification with KAPA HiFi enzyme.
- Compatibility with a wide range of DNA sample types, inputs and sequencing applications, and flexibility with respect to fragment size, adapter design, and library amplification.
- Qualified automation methods.
KAPA HyperPlus Guide to Success
Spatial genomic heterogeneity within localized, multifocal prostate cancer
Library Amplification
Library preparation for next-generation sequencing (NGS) often involves an amplification step for obtaining fragments enriched for adapter ligated ends and for increasing the library size, both of which are essential for effective detection of sequences during a sequencing run. However, PCR can often introduce bias for sample composition, leading to potential underrepresentation of these fragments in a sequencing read.
Roche Sequencing Solutions offers KAPA Library Amplification Kits, which contain KAPA HiFi DNA polymerase, a novel enzyme engineered through our directed evolution technology, for ultra-high fidelity and robustness. KAPA HiFi improves the overall sequencing quality with low amplification bias, more uniform coverage of difficult regions, lower duplication rates compared to other options, and amplifies NGS libraries with industry-leading fidelity.
KAPA Library Amplification Kits include KAPA HiFi DNA Polymerase, a novel enzyme engineered using our directed evolution technology for ultra-high fidelity and robustness. KAPA HiFi has become the enzyme of choice for next-generation sequencing (NGS) library amplification due to its ability to amplify complex DNA populations with high fidelity, high efficiency and very low amplification bias. This results in lower duplication rates and improved coverage of GC- and AT-rich regions, promoters, low-complexity and other challenging regions.
Benefits
- Low amplification bias.
- Improved sequencing coverage uniformity of difficult regions.
- Industry-leading fidelity.
- Fewer cycles to achieve equivalent yields due to higher amplification efficiency.
KAPA HiFi Uracil+ DNA Polymerase is a modified version of KAPA HiFi DNA Polymerase that is engineered to tolerate uracil residues in bisulfite-treated DNA. Bisulfite treatment of DNA results in the conversion of unmethylated cytosine residues into uracil, while the methylated residues are left unmodified. Enzymes used for library amplification in bisulfite-sequencing workflows must be able to read through uracil residues and tolerate low concentrations of AT-rich DNA. Traditional high-fidelity DNA polymerases are typically not suitable for this type of application as the enzymes stall when a uracil is encountered. KAPA HiFi Uracil+ DNA Polymerase has been developed to read through uracil residues while still retaining the performance benefits of HiFi DNA Polymerase, such as high yields, low-bias, and uniform sequencing coverage.
Benefits
- Higher yields and reduced size bias for templates containing uracil compared to other methods.
- More uniform bisulfite-sequencing coverage and improved representation of AT-rich sequences compared to competitor kits.
- Effective amplification through uracil residues.
- Reduced cycles and lower PCR duplicates through high-efficiency amplification
- Improved representation of all library fragments and sequence regions due to low amplification bias
- Greater tolerance to AT-rich regions
- Improved bisulfite-sequencing read quality
Targeted DNA demethylation and activation of endogenous genes using programmable TALE-TET1 fusion proteins
RNA Library Preparation
RNA sequencing (RNA-Seq) leverages next generation sequencing (NGS) to provide insights into global transcriptional events, coding mRNA transcripts or on the profiles of RNA species, such as lncRNA. RNA library preparation for NGS utilizes specific strategies, such as enrichment of mRNA or depletion of rRNA, based on the objective of the sequencing experiment.
Roche Sequencing Solutions offers a portfolio of products containing high-quality enzymes developed through our Directed Evolution Technology for constructing RNA libraries with minimal GC bias and even sequence coverage. KAPA RNA Library Preparation Kits include KAPA HiFi DNA Polymerase, which provides industry-leading fidelity. Roche Sequencing Solutions provides a complete library preparation solution with KAPA Adapters and KAPA Pure Beads. These kits are designed to be automation-friendly on a majority of commercially available liquid handlers.
The KAPA RNA HyperPrep Kits utilize a novel chemistry that enables the combination of enzymatic steps and fewer reaction purifications, resulting in a truly streamlined solution for the preparation of high-quality RNA-seq libraries. The strand-specific workflow is flexible, supporting library construction from lower-input amounts and degraded samples and is compatible with mRNA capture and ribosomal depletion. KAPA RNA HyperPrep Kits contain all reagents required for RNA library preparation, with the exception of KAPA Adapters (available separately).
Benefits
- Reduced hands-on and overall time through fewer enzymatic and reaction cleanup steps.
- Strand-specific, sequencing-ready libraries from input RNA in ~4 hours.
- Single-day library construction, including mRNA capture and rRNA depletion with the KAPA RNA HyperPrep workflow.
- Robust performance across different sample types and input amounts.
- High throughput and consistency with an automation-friendly workflow.
KAPA mRNA HyperPrep Kits enrich mRNA in samples and provide a focused view of the protein-coding regions in the transcriptome. mRNA capture beads are used prior to library preparation with the KAPA RNA HyperPrep workflow. The kits enrich for mRNA over non-polyadenylated species such as ribosomal, precursor and noncoding RNAs.
Benefits
- mRNA capture in ~1.5 hours.
- Robust performance across different sample types and input amounts.
- High success rates with low-input samples.
- Qualified automation methods.
- Waste fewer reads due to rRNA carryover and PCR duplicates.
- Identify more unique transcripts and genes with equivalent sequencing.
The KAPA RNA HyperPrep Kit with RiboErase (HMR) targets and depletes rRNA using DNA probes and RNase H to provide improved coverage of transcripts of interest, including precursor mRNAs and important regulatory species, such as noncoding RNAs. Sequencing of rRNA-depleted total RNA samples provides a more comprehensive representation of the whole transcriptome. This kit is suitable for total RNA input from human, mouse and rat species only.
Benefits
- rRNA depletion and purification in ~2.5 hours.
- Robust performance across different sample types and input amounts.
- High success rates with low-input and degraded samples.
- Qualified automation methods.
- Waste fewer reads on rRNA carryover and PCR duplicates.
- Identify more unique transcripts and genes with equivalent sequencing.
The KAPA Stranded RNA-Seq Kit combines the use of a "with bead" protocol with KAPA HiFi DNA Polymerase selected through our directed evolution technology for high efficiency, industry-leading fidelity and low-bias amplification to provide superior quality RNA libraries with flexible input amounts. Roche Sequencing Solutions provides a complete library preparation solution with KAPA Adapters and KAPA Pure Beads, available separately.
Benefits
- Tunable fragmentation for various sequencing applications.
- High yield of adapter-ligated library.
- High coverage of GC-rich and low-abundance transcripts.
- Qualified automation methods.
- Comparable transcript coverage at various inputs.
- Reproducible and uniform distribution of reads across the transcript.
The KAPA Stranded mRNA-Seq Kit makes use of KAPA mRNA Capture Beads prior to library preparation to enrich for mRNA over non-polyadenylated species such as ribosomal, precursor or noncoding RNAs. The kits contain KAPA HiFi DNA Polymerase, selected through our directed evolution technology for high-efficiency and low-bias library amplification and are optimized for the improved coverage of GC-rich and low-abundance transcripts. Roche Sequencing Solutions provides a complete library preparation solution with KAPA Adapters and KAPA Pure Beads, available separately.
Benefits
- Focused sequencing of protein-coding transcripts.
- Higher coverage of GC-rich transcripts compared to other options.
- Identification of more transcripts and genes compared to other options.
- Qualified automation methods.
The progesterone antagonist mifepristone/RU486 blocks the negative effect on life span caused by mating in female Drosophila
The KAPA Stranded RNA-Seq Kit with RiboErase (HMR) targets and depletes rRNA from various sample types (human, mouse, rat) and provides more consistent and effective rRNA depletion than traditional bead-based capture methods. Efficient rRNA depletion prior to library preparation results in increased coverage of transcripts of interest, including noncoding and precursor transcripts. This kit is suitable for total RNA input from human, mouse and rat species only. Roche Sequencing Solutions provides a complete library preparation solution with KAPA Adapters and KAPA Pure Beads, available separately.
Benefits
- Up to 99.98% rRNA depletion from various sample types.
- Compatible with degraded inputs, including FFPE.
- High coverage of GC-rich and low-abundance transcripts.
- Qualified automation methods.
- Superior rRNA depletion with low inter-sample variability.
- Highly efficient and reproducible rRNA depletion with degraded inputs, such as FFPE.
- Maximized sequencing capacity by reducing rRNA reads.
Library Preparation Accessories
Library preparation for next generation sequencing (NGS) involves several steps – fragmentation of nucleic acids to required lengths, end repair and ligation of adapters to ends of target sequences, library amplification and quantification. A critical step in DNA and RNA library construction is to select for DNA fragments of appropriate size and to remove adapter dimers. Adapter quality and bead performance can have a profound impact on the outcome of library construction, particularly when working with low-input and challenging samples. Roche Sequencing Solutions offers high-quality KAPA Pure Beads and KAPA Adapters that are functionally tested for NGS library construction. They are supplied as stand-alone products to support flexibility.
KAPA Pure Beads offer a tunable and highly consistent solution for reaction purification and size selection in DNA and RNA next-generation sequencing (NGS) library construction workflows.
Benefits:
- High recovery of single- and double-stranded DNA (1 ng – 5 μg) in a single cleanup.
- Fast and efficient cleanups to remove unwanted reaction components.
- Easy substitution into bead-based workflows.
- Adjustable size selection.
- Automation friendly.
Compatible with all KAPA library preparation kits for DNA and RNA sequencing, the high-quality KAPA Single-Indexed and Dual-Indexed Adapters can be used with Illumina library construction protocols employing standard adapters. Compatible with the HyperCap Workflow v2.0.
KAPA Library Quantification Kits contain all the reagents needed for the accurate, reliable and reproducible qPCR-based quantification of next-generation sequencing (NGS) libraries prepared for sequencing on Illumina and IonTorrent platforms. Kits include KAPA SYBR FAST qPCR Master Mix (formulated with different passive reference dyes for different qPCR instruments), a platform-specific library quantification primer premix, and a pre-diluted set of DNA standards.
Preparation of a DNA library for next generation sequencing (NGS) platforms entails several steps that have a direct influence on the quality of the sequencing library, and ultimately on the reliability of sequencing results. Roche Sequencing Solutions offers best-in-class* DNA library preparation kits containing high-quality enzymes and reaction buffers developed through our Directed Evolution Technology. The KAPA Library Preparation Kits include modules required for end repair, A-tailing, ligation, and amplification (for Illumina platforms) and offer complete library preparation solutions with KAPA Adapters and KAPA Pure Beads.
Library preparation for next-generation sequencing (NGS) often involves an amplification step for obtaining fragments enriched for adapter ligated ends and for increasing the library size, both of which are essential for effective detection of sequences during a sequencing run. However, PCR can often introduce bias for sample composition, leading to potential underrepresentation of these fragments in a sequencing read.
RNA sequencing (RNA-Seq) leverages next generation sequencing (NGS) to provide insights into global transcriptional events, coding mRNA transcripts or on the profiles of RNA species, such as lncRNA. RNA library preparation for NGS utilizes specific strategies, such as enrichment of mRNA or depletion of rRNA, based on the objective of the sequencing experiment.
Library preparation for next generation sequencing (NGS) involves several steps – fragmentation of nucleic acids to required lengths, end repair and ligation of adapters to ends of target sequences, library amplification and quantification. A critical step in DNA and RNA library construction is to select for DNA fragments of appropriate size and to remove adapter dimers. Adapter quality and bead performance can have a profound impact on the outcome of library construction, particularly when working with low-input and challenging samples. Roche Sequencing Solutions offers high-quality KAPA Pure Beads and KAPA Adapters that are functionally tested for NGS library construction. They are supplied as stand-alone products to support flexibility.