The seven steps of sample prep

Converting a sample to a sequencing library typically involves seven steps, and every step of the process has the potential to reduce or bias the value. To preserve sample integrity and convert nucleic acids into sequenceable molecules with minimal loss, you have to work with the best sample prep solutions for each step.

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Next-generation sequencing
The seven steps of your workflow.

Roche Sample Prep Solutions offer integrated service and support for your next-generation sequencing (NGS) workflow, from sample collection to sequencing ready libraries.

Step 1: Sample collection

The Cell-Free DNA Collection Tube is a direct-draw tube for the collection, stabilization and transportation of whole blood specimens. It effectively prevents blood coagulation and cell lysis, and it preserves nucleated cells to enable efficient analysis of cell-free DNA (cfDNA).

Step 2: Sample enrichment

The AVENIO Millisect System is an automated, high-performance tissue dissection system that enables precise and consistent recovery of formalin-fixed paraffin-embedded (FFPE) tissue for molecular pathology. The system is optimized to fit easily into a variety of clinical workflow configurations. It improves diagnostic testing results by reducing false negatives, and can help you efficiently extract more clinically relevant information from every sample.

Step 3: Nucleic acid extraction

The Roche family of MagNA Pure Systems automates and simplifies nucleic acid extraction, which dramatically reduces the handling errors associated with manual techniques. The MagNA Pure Systems have been optimized for NGS workflows using new protocols compatible with Roche Sample Prep Solutions, bringing you confidence in your sequencing results.

Step 4: Sample quantification/QC

KAPA hgDNA Quantification and QC Kits contain all the consumables needed for the qPCR-based quantification and quality assessment of human genomic DNA samples prior to NGS library construction. Each kit contains the KAPA SYBR FAST qPCR Master Mix, optimized for high-performance SYBR Green I-based qPCR through our Directed Evolution Technology. A set of pre-diluted DNA standards and primer premixes targeting different portions of a highly conserved single-copy human locus. Achieve absolute quantification: the primer pair defines the shortest fragment and the additional primers provide information about the amount of amplifiable template in the DNA sample. Quality scores (or Q-ratios) generated with the kit may be used to predict the outcome of library construction or to tailor workflows for samples of variable quality, particularly DNA.

Step 5: Library preparation

KAPA HyperPrep Kits offer a streamlined library preparation protocol, combining several enzymatic steps and eliminating bead cleanups to 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 solutions with KAPA Adapters and KAPA Pure Beads.

KAPA HyperPlus Kits, our most advanced library preparation option, provide a streamlined workflow that includes fully automatable fragmentation and library preparation in a single tube. This integrated solution combines enzymatic fragmentation, with the quality of mechanical shearing, and the speed and convenience of tagmentation-based workflows. KAPA HyperPlus Kitsoffer a 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.

The KAPA RNA HyperPrep Kits utilize a novel chemistry to enable the combination of enzymatic steps and fewer reaction purifications. This results 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. It 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).

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 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.

KAPA HiFi Uracil+ DNA Polymerase is a modified version of KAPA HiFi DNA Polymerase. It 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. 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

Step 6: Target enrichment and capture

SeqCap® EZ Probes use an innovative single-tube solution-based hybridization capture technology that enables enrichment of any genomic region of interest. They are built upon an optimized design algorithm and manufactured with high probe density and demonstrate uniform and higher depth of coverage.

KAPA Dual-Indexed Adapters enable greater multiplexibility compared to existing options by utilizing up to 96 sample barcodes. Roche Universal Blocking Oligo Kits eliminate the need for adapter-matched blocking oligos, affording a simple and faster workflow.

HyperCap products provide an integrated next-generation sequencing (NGS) sample preparation solution that combines all reagents and accessories required for library preparation and target enrichment in a single workflow. The HyperCap Workflow v2.0 integrates high-efficiency KAPA Library Preparation Kits and KAPA Dual-Indexed Adapters with performance-optimized SeqCap® EZ probes and Roche Universal Blocking Oligo (UBO) Kit. The HyperCap Workflow v2.0 is an improved and streamlined version of our SeqCap EZ HyperCap Workflow. It is automation friendly and offers the convenience of ordering and support from a single vendor.

Step 7: Library quantification/QC

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 and SeqCap are for Research Use Only. Not for use in diagnostic procedures. Millisect and MAGNA PURE 24 and 96 products are for use for in vitro diagnostics Cell-Free DNA Collection Tubes are available both in RUO* and CE-IVD** formats.

*For Research Use Only. Not for use in diagnostic procedures.
**Available for countries that accept CE-Mark.]

Talk to a local expert

We offer proven, simple and complete solutions to optimise your sequencing resources. How can this help you enhance sample prep? We would love to discuss your options. Contact your Roche representative today, and we’ll be in touch.

Roche Sample Prep Solutions

From sample collection to library quantification, our integrated workflow solutions for different sample types and sequencing applications are proven, complete and simple.

Proven

Our portfolio of high-quality sample prep products have served the life sciences, translational and clinical research community for over a decade.

Simple

Streamlining every step of the process, from pipetting to purchasing, improves sequencing success and frees up your time and resources.

Complete

Qualified, complete workflow solutions for a wide range of sample types and applications give you greater peace of mind.

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