KAPA RNA HyperPrep Kits

Rapid, robust, and reliable

KAPA RNA HyperPrep Kits

KAPA RNA HyperPrep Kits

These fast, robust RNA library preparation kit can be used with a wide variety of RNA input types, qualities, and amounts, and they also enable 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

 

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

 

KAPA RNA HyperPrep Kits 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

 

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).

Additional Resources

Additional resources for KAPA RNA HyperPrep Kits are available

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