KAPA RNA HyperPrep Kits

For Research Use Only. Not for use in diagnostic procedures.
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KAPA RNA HyperPrep Kits for RNA-seq libraries

KAPA RNA HyperPrep Kits are a comprehensive, modular solution for a variety of RNA sequencing applications, from mRNA capture and rRNA depletion to whole transcriptome sequencing (WTS) and custom transcript depletion. Each kit includes KAPA Pure Beads and KAPA HiFi Polymerase ReadyMix, ensuring optimal performance and high-quality results.

These kits, with their rapid workflows, stand out amid the rest for their:

  • Ease of use, with simple protocols that require under 2 hours of total hands-on time;
  • Delivery of superior results, including higher yields of unique variants with fewer amplification cycles; 
  • High efficiency in variant detection, reducing the number of sequencing runs needed to achieve necessary coverage depth.

By choosing Roche, you gain more than just high-quality kits—you also gain the support of our dedicated field scientists, expert applications development team, and automation specialists. 

Benefits of KAPA RNA HyperPrep Kits
Diagram showing advantages of KAPA RNA HyperPrep, including performance, rapidity, automatability, and support
Photo of automated NGS library preparation

Thinking of automating library prep on your existing liquid handler?

Roche’s NGS Automation Support Team, in collaboration with non-Roche liquid handling vendors, creates menus of automated methods for KAPA library prep reagents.

Let’s talk about the next steps to get you up and running with automating your NGS library prep protocols. Visit support for non-Roche liquid handlers to learn more.

Diagram of workflows for RNA-seq with ribodepletion and with mRNA capture
kapa rna hyperprep

Depletion of rRNA, user-defined RNA, and globin transcript.

Whole-transcriptome sequencing provides a complete view of the transcriptome, including immature and noncoding RNAs. In these workflows, it is important to remove rRNA (up to 95% of the total RNA sample) prior to library prep in order to minimize the number of wasted sequencing reads. This method is effective with degraded RNA inputs not suitable for mRNA capture. Enzymatic depletion can also be used to remove other transcripts, such as globin mRNA and/or other highly abundant RNAs.

mRNA Capture

mRNA capture sequencing is used for the analysis of coding transcripts in eukaryotic samples, allowing researchers to detect and quantify relative expression levels. This workflow enriches for mRNA over non-polyadenylated species such as ribosomal, precursor, and non-coding RNAs.

KAPA RNA HyperPrep Kits provide streamlined and strand-specific library construction.

The novel chemistry employed in KAPA RNA HyperPrep Kits allows for fewer and shorter enzymatic steps, reducing hands-on time and overall library prep time. rRNA depletion with KAPA RiboErase (HMR) or KAPA RiboErase (HMR) Globin Kits adds approximately 2.5 hours to the overall workflow time, whereas mRNA capture adds approximately 1.5 hours. The entire workflow, from input RNA to sequencing-ready library, can easily be completed in a standard workday. All KAPA RNA HyperPrep library construction workflows are automation friendly.

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KAPA RNA HyperPrep Kits Specifications and Comparison

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KAPA RNA HyperPrep Kits Specifications and Comparison

 

Product Name KAPA RNA HyperPrep Kits KAPA mRNA HyperPrep Kits KAPA RNA HyperPrep Kits with RiboErase (HMR) KAPA RNA HyperPrep Kits with RiboErase (HMR) Globin
Library Preparation Duration 4 hours 5.5 hours 6.5 hours 6.5 hours
Sample Input Amount 1-100 ng into library prep 50 ng - 1 µg into mRNA capture 25 ng - 1 µg into rRNA depletion 25 ng - 1 µg into rRNA depletion
Sample Types

High-quality total RNA


Degraded or FFPE total RNA


Poly(A)-selected and/or depleted RNA

 High-quality total RNA

High-quality total RNA


Degraded or FFPE total RNA

High-quality total RNA


Degraded or FFPE total RNA


Whole blood total RNA
Species

Eukaryotic (Animal, Plant, etc.)


Prokaryotic (Bacteria, etc.)


Viral

 Eukaryotic (Animal, Plant, etc.)

Human, Mouse, and Rat


Other species can be supported via custom rRNA depletion

Human, Mouse, and Rat


Other species can be supported via custom rRNA depletion
Stranded RNA-sequencing Yes Yes Yes Yes
Amplification

KAPA HiFi DNA Polymerase


Compatible with low bias amplification

KAPA HiFi DNA Polymerase


Compatible with low bias amplification

KAPA HiFi DNA Polymerase


Compatible with low bias amplification

KAPA HiFi DNA Polymerase


Compatible with low bias amplification
Reaction Purification KAPA Pure Beads included KAPA Pure Beads included KAPA Pure Beads included KAPA Pure Beads included
Multiplexing Yes.  KAPA Adapters are not supplied, but available separately Yes.  KAPA Adapters are not supplied, but available separately Yes.  KAPA Adapters are not supplied, but available separately Yes.  KAPA Adapters are not supplied, but available separately
Applications

Whole-transcriptome sequencing


Gene expression with target enrichment


Splice junction and gene fusions with target enrichment

Gene expression


Annotation


Splice junction and gene fusions

Whole-transcriptome


Long non-coding RNA Annotation


Gene expression


Splice junction and gene fusions

Whole-transcriptome


Long non-coding RNA Annotation


Gene expression


Splice junction and gene fusions
Automation Friendly Yes.  Demonstrated and verified on
many liquid handling systems.
Learn more		 Yes.  Demonstrated and verified on
many liquid handling systems.
Learn more		 Yes.  Demonstrated and verified on many
liquid handling systems.
Learn more		 Yes.  Demonstrated and verified on many
liquid handling systems.
Learn more
Compatible Sequencing Platforms Illumina Sequencers Illumina Sequencers Illumina Sequencers Illumina Sequencers
Storage Information

Enzymes and buffers can be stored for up to 10 months at -20°C

 

KAPA Pure Beads can be stored for up to 10 months at 4°C

Enzymes and buffers can be stored for up to 10 months at -20°C

 

KAPA Pure Beads can be stored for up to 10 months at 4°C

Enzymes and buffers can be stored for up to 10 months at -20°C

 

KAPA Pure Beads can be stored for up to 10 months at 4°C

Enzymes and buffers can be stored for up to 10 months at -20°C

 

KAPA Pure Beads can be stored for up to 10 months at 4°C
The first-generation KAPA Stranded RNA-seq Kits are still available for purchase. For more information about these older RNA library kits, please click here.
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KAPA RNA HyperPrep Kits

These fast, robust RNA library preparation kits can be used with a wide variety of RNA input types, qualities, and amounts. They enable many different workflow options, including ribosomal depletion and mRNA capture for eukaryotic or prokaryotic RNA-seq libraries.

KAPA RNA HyperPrep with RiboErase (HMR)

  • Includes all reagents for efficient enzymatic depletion of ribosomal RNA (rRNA) from human, mouse, and rat (HMR) samples of high-or low-quality RNA
  • Provides a more comprehensive view of the whole transcriptome compared to mRNA capture
  • Can be used for custom depletion of rRNA from other organisms when the HMR oligos are replaced with custom sequences
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Figure 1. Efficient rRNA depletion

Figure 2. Efficient rRNA depletion
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Efficient rRNA depletion using KAPA RNA HyperPrep with RiboErase (HMR) leads to fewer non-informative reads and the detection of more unique transcripts compared to other suppliers.

Libraries were generated in quadruplicate using variable inputs of Universal Human Reference (UHR) RNA (Agilent Technologies) with rRNA de[pletion prior to library construction. Where present, error bars represent the standard deviation. For 25 ng samples, paired end (2 x 100 bp) sequencing was performed using an Illumina® HiSeq® 2500 instrument. Reads aligning to rRNA were removed, and reads were randomly subsampled to 14 M for comparative analyses. Transcripts were quantified using RNA-SeQC. The 10 ng input amount is lower than the validated minimum input for both KAPA RNA HyperPrep workflows; for these samples, paired-end (2 x 75 bp) sequencing was performed using an Illumina NextSeq 500 instrument. Reads were randomly subsampled to 14 M for comparative analysis prior to removing reads aligning to rRNA and subsequent marking of duplicates. Transcripts were quantified using Kallisto (data not plotted due to analysis and sequencing depth differences).

KAPA RNA HyperPrep with RiboErase (HMR) Globin

  • Includes all reagents for efficient enzymatic depletion of globin mRNA from blood-derived RNA samples from high- or low-quality RNA
  • Includes reagents for the depletion of human, mouse, or rat (HMR) rRNA
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Fig 3. KAPA RNA HyperPrep Kits

Fig 4. KAPA RNA HyperPrep Kits

Fig 5. KAPA RNA HyperPrep Kits
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KAPA RNA HyperPrep Kit with RiboErase (HMR) Globin simultaneously removes rRNA and globin mRNA from blood-derived RNA. Together, effective RNAase H depletion and highly efficient library construction result in fewer non-informative reads 

(A) compared to the workflow from Supplier I, which employs bead-based depletion (orange). This translates to more complex libraries (B) and a larger number of unique transcripts detected (C).

Libraries were prepared from different inputs of RNA extracted from human blood, as indicated on the x-axis of each graph. The 25 ng input is lower than the recommended minimum input for the Supplier I workflow. Paired-end (2 x 125 bp) sequencing was performed on an Illumina® HiSeq® 2500 instrument. Data were sub-sampled to 17 M reads per sample for analysis. Each bar represents the average of three technical replicates. Transcript abundance was quantified using Kallisto. To assess off-target depletion, transcript abundances were aggregated at the gene level and TMM-normalized prior to differential expression analysis. The expression profiles of libraries generated with or without globin depletion were compared to assess off-target depletion for each workflow.

KAPA mRNA HyperPrep Kit

  • Includes reagents for the enrichment of eukaryotic mRNA from high-quality total RNA
  • Provides a focused view of the protein-coding regions of the transcriptome
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Fig 6

figure 7
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Efficient mRNA capture using KAPA mRNA HyperPrep leads to fewer non-informative reads and the detection of more unique transcripts compared to other suppliers. 

Libraries were generated in quadruplicate using variable inputs of Universal Human Reference (UHR) RNA (Agilent Technologies) with mRNA capture prior to library construction. For 50 ng samples, paired end (2 x 100 bp) sequencing was performed using an Illumina® HiSeq® 2500 instrument. Reads aligning to rRNA were removed, and reads were randomly subsampled to 14 M for comparative analyses. Transcripts were quantified using RNA-SeQC. The 10 ng input amount is lower than the validated minimum input for both KAPA RNA HyperPrep workflows; for these samples, paired-end (2 x 75 bp) sequencing was performed using an Illumina NextSeq 500 instrument. Reads were randomly subsampled to 14 M for comparative analysis prior to removing reads aligning to rRNA and subsequent marking of duplicates. Transcripts were quantified using Kallisto (data not plotted due to analysis and sequencing depth differences).

Ask a Scientist: How does RNA sequencing work?

How do I sequence RNA, and what can I learn from RNA-seq data? This video describes the process of RNA sequencing, including preparation of RNA-seq libraries for NGS. It also explains how RNA-seq data can provide insights into cellular activity, including differential gene expression, splice variants, and expression of novel transcripts. Presented by Dustin Masser, Field Application Specialist at Roche Sequencing and Life Science

Importance of performing ribosomal RNA depletion

In this short video (~3 min), you’ll learn how the depletion of ribosomal RNA (rRNA) can improve your RNA-seq results. You’ll also learn about the differences between enzymatic and bead-based methods, and how these methods can be modified to enable custom depletion of user-defined transcripts. Presented by Nicholas Hapshe, Roche Sequencing and Life Science.

What is Stranded RNA Sequencing?

What is stranded RNA sequencing, and how is it different from other RNA-seq workflows? This video describes the library preparation workflow for stranded RNA-seq, and explains the benefits of using this method. These include the ability to separately identify transcripts originating from each DNA strand, increasing the accuracy of RNA-seq data and providing additional insights-- especially important when genes on opposite strands overlap.

Presented by Dustin Masser, Roche Sequencing and Life Science

Webinars

RNA:Seq: Addressing the Challenges using KAPA RNA HyperPrep

RNA-sequencing (RNA-seq) and the study of transcriptomes have become essential in both molecular and clinical research. Many researchers and sequencing service providers have found RNA-seq difficult to adopt due to various challenges. 

In this webinar, we discuss:

  •  What RNA-seq is and why it is crucial in research
  • Unique RNA-seq challenges in sample handling, library preparation, and data analysis
  •  KAPA RNA HyperPrep workflows and how they can help overcome these challenges
  • Q&A with an experienced Roche field application specialist

Presented by Drew Cheney, Field Applications Scientist, Roche Sequencing and Life Science

Automating RNA-seq Library Preparation

Automating RNA-sequencing (RNA-seq) library preparation offers advantages such as higher sample throughput, less hands-on time, fewer human errors, greater reproducibility, and better process control and workflow standardization. However, developing and implementing an automated RNA-seq library prep protocol on a liquid handling platform can be challenging and time-consuming.

In this webinar, we discuss the automation of RNA-seq library prep, including:

  • Benefits of automating RNA-seq library prep
  • Tips and tricks of NGS automation
  • Overview of the Roche Sequencing automation program
  • Examples of automated workflows for KAPA RNA HyperPrep
RNA-seq Applications for SARS-CoV-2 Research

In this presentation, you’ll learn about high-throughput SARS-CoV-2 research using RNA-seq. Highlighted methods include whole-transcriptome and target-enriched sequencing, using both hybridization-based and amplicon workflows. Presented by Sarah Trusiak, Senior Applications Scientist at Roche Sequencing and Life Science on August 28, 2020.

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Realted products

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    KAPA Cat. No
    		 Roche Cat. No
    		 Description
    		 Kit Size
    KK8560
    		 08098131702

    KAPA RNA HyperPrep with RiboErase (HMR)

    		 24 libraries
    KK8561
    		 08098140702

    KAPA RNA HyperPrep with RiboErase (HMR)

    		 96 libraries
    KK8540
    		 08098093702
    		 KAPA RNA HyperPrep Kit 24 libraries
    KK8541
    		 08098107702
    		 KAPA RNA HyperPrep Kit 96 libraries
    KK8580
    		 08098115702
    		 KAPA mRNA HyperPrep Kit 24 libraries
    KK8581
    		 08098123702
    		 KAPA mRNA HyperPrep Kit 96 libraries
    KK8562
    		 08308314702

    KAPA RNA HyperPrep Kit with RiboErase (HMR) Globin

    		 24 libraries
    KK8563
    		 08308241702

    KAPA RNA HyperPrep Kit with RiboErase (HMR) Globin

    		 96 libraries
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    KAPA Cat. No
    		 Roche Cat. No
    		 Description
    		 Kit Size
    KK8721
    		 08278539001
    		 KAPA Adapter Dilution Buffer (25 mL) 25 mL
    KK8727*
    		 08861919702

    KAPA Unique Dual-Indexed Adapter Kit, (15 µM)

    		 96 adapters x 20 µl each
    *Contains KAPA Adapter Dilution Buffer, as well as three additional sealing films to support multiple use
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