Kapa - Library Preparation

Preparing target nucleic acids for next-generation sequencing

Library construction is a critical step in the next generation sequencing (NGS) workflow. DNA or RNA samples are first fragmented to the appropriate size and then sequencing platform-specific adapters are added. In most cases, libraries have to be amplified before the next step in the process. Each step can potentially introduce errors or biases that could significantly impact library quality and ultimately affect the quality and reliability of sequencing results. It is therefore imperative to use the highest quality reagents and optimized protocols to obtain the best sequencing libraries, lowest bias, and the highest and most uniform coverage.

Product List




Library Quantification

subcat_libquant_banner_1880x560.pngKAPA 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.

Links



DNA Library Preparation

subcat_dnalibprep_banner_1880x560.pngPreparation 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

KAPA HyperPrep Kits

KAPA HTP/LTP Library Preparation Kits



Library Amplification

subcat_lak_banner_1880x560_right.pngLibrary 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

KAPA HiFi Uracil+ Kits



RNA Library Preparation

subcat_rnalibprep_banner_1880x560.pngRNA 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.

KAPA RNA HyperPrep Kits

KAPA mRNA HyperPrep Kits

KAPA RNA HyperPrep Kits with RiboErase

KAPA Stranded RNA-Seq Kits

KAPA Stranded mRNA-Seq Kits

KAPA Stranded RNA-Seq Kits with RiboErase



Library Preparation Accessories

subcat_libprepaccessories_ban_1880x560_right.pngLibrary 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

KAPA Adapters

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