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.

Benefits of KAPA RNA Library Preparation Kits

  • High coverage uniformity and coverage of GC-rich regions and low-abundance transcripts
  • Low duplication rates resulting in increased sequencing throughput and low bias
  • Ability to automate
  • Ability to enrich for specific species of RNA
Selection Chart
  No RNA Enrichment
mRNA Capture
Ribosomal Depletion
cDNA Library Prep Workflow
Standard
Streamlined
Standard
Streamlined
Standard
Streamlined
KAPA Product
KAPA Stranded RNA-Seq Kit
KAPA RNA HyperPrep Kit
KAPA Stranded mRNA-Seq Kit
KAPA mRNA HyperPrep Kit
KAPA Stranded RNA-Seq Kit with RiboErase
KAPA RNA HyperPrep Kit with RiboErase
Library Prep Workflow time
6 – 8 hr
4 hr
8 – 10 hr
5.5 hr
10 – 12 hr
6.5 hr
Input Amount
10 – 400 ng into library prep
1 – 100 ng into library prep
100 ng – 4 μg into mRNA capture
50 ng – 1 μg into mRNA capture
100 ng – 1 μg into rRNA depletion
25 ng – 1 μg into rRNA depletion
Sample Type
High-quality Total RNA High-quality Total RNA High-quality Total RNA
Degraded or FFPE Total RNA   Degraded or FFPE Total RNA
Enriched RNA
   
Species
Eukaryotic (Animal, Plant, etc.)
Eukaryotic (Animal, Plant, etc.)
Human, Mouse, and Rat
Prokaryotic (Bacterial, etc.)
   

Differentiating Applications

Targeted RNA-Seq
mRNA-Seq
Non-coding RNA
Whole Transcriptome
  Whole Transcriptome
Shared Applications
Gene Expression Analysis; Detection of Gene Fusions, Isoforms, and other Structural Variants; Novel Transcript Identification; SNV Discovery
Strand-specific
Yes
Automation friendly
Yes
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KAPA RNA HyperPrep Kits

As the latest generation of library preparation kits, KAPA RNA HyperPrep Kits utilize a novel chemistry and combines enzymatic steps to achieve fewer reaction purifications and yield high-quality RNA-Seq libraries in a single day.

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KAPA mRNA HyperPrep Kits

KAPA mRNA HyperPrep Kits enrich for polyadenylated species and provide a focused view of the coding regions. Kits provide high coverage uniformity and minimal PCR duplicates.

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KAPA RNA HyperPrep Kits with RiboErase

KAPA RNA HyperPrep Kits with RiboErase provide consistent and effective rRNA depletion and reduce the number of reads wasted due to alignment to rRNA loci. Kits provide high coverage uniformity and yield high-quality libraries with degraded inputs, including FFPE samples.

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KAPA Stranded RNA-Seq Kits

As the first generation RNA library preparation kits, the KAPA Stranded RNA-Seq Kits combine a “with-bead” protocol with the robustness of KAPA HiFi DNA Polymerase to yield high-quality RNA libraries with high sequence coverage and low bias.

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KAPA Stranded mRNA-Seq Kits

KAPA Stranded mRNA-Seq Kits make use of KAPA mRNA Capture Beads prior to library preparation and provide a focused sequencing of protein-coding transcripts with high coverage of GC-rich and low-abundance transcripts.

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KAPA Stranded RNA-Seq Kits with RiboErase

KAPA Stranded RNA-Seq Kits with RiboErase provide consistent and effective rRNA depletion compared to traditional bead-capture methods* yielding high coverage of transcripts of interest, including noncoding and precursor transcripts. Compatible with degraded inputs, including FFPE samples.

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Data on file.