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 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.
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.
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 |
Compatible with low bias amplification |
Compatible with low bias amplification |
Compatible with low bias amplification |
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 |
Genialis™ Data Analysis and Visualization Solution | Yes Learn more |
Yes Learn more |
Yes Learn more |
Yes Learn more |
KAPA RiboDesigner Custom rRNA Depletion Support | N/A | N/A | Yes Learn more |
Yes 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 |