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Kapa biosystems/KAPA Stranded RNASeq with RiboErase/KK8000/5 mL

Description:

KAPA Pure Beads (5 mL)

KAPA Stranded RNA-Seq with RiboErase

Evolved to focus.

The KAPA Stranded RNA-Seq Kit with RiboErase offers a high-quality, comprehensive solution for transcriptome sequencing. By utilizing a targeted enzymatic method for depletion, our workflow enables superior reduction of ribosomal RNA (rRNA) and a more complete representation of the transcriptome, including precursor mRNAs and non-coding RNA (ncRNA). Kits also contain KAPA HiFi for high-efficiency and low-bias library amplification, and include a streamlined, “with-bead” protocol.

  • Up to 99.98% rRNA depletion*
  • Flexible input of 100 ng – 1 μg total RNA from human, mouse, or rat species*
  • Robust and reproducible across various input amounts
  • Automation-friendly workflow

NEW! KAPA Dual-Indexed Adapter Kits are now available. For more information on KAPA Adapter Kits, scroll down to the Ordering section, or download the KAPA Adapter and Bead Calculator.

Download our Adapter and Bead Calculator

 

*Data on file.
For Research Use Only. Not for use in diagnostic procedures.

Product Highlights

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Industry leading rRNA depletion

  • Superior rRNA depletion from high-quality and FFPE samples*
  • More economical NGS sequencing due to decreased rRNA reads, providing deeper sequencing of transcripts of interest
High-quality sequencing data obtained from KAPA Stranded RNA-Seq Kit with RiboErase when compared to the equivalent Illumina or NEB kits. With equivalent mapping rates, KAPA Stranded RNA-Seq Kit with RiboErase provides lower duplication rates, better coverage uniformity and consistently detects more unique transcripts and genes. This enables a more sensitive detection of the expressed genes within the sample. Data on file.
Improved coverage of GC-rich transcripts. IGV image showing coverage and splice junction tracks of the haemoglobin alpha transcript (HBA1) which has a GC content of 62.8%. The GC content track shows GC-rich regions in red, and AT-rich regions in blue. More even coverage, improved splice-site recognition, and more representative plot compared to the undepleted sample (red) is seen when libraries are prepared with the KAPA Stranded RNA-Seq Kit with RiboErase (green) compared to the equivalent Illumina (orange) or NEB (blue) kits. Data on file.
Better detection of low-abundance transcripts. IGV image showing coverage and splice junction tracks of MALAT1, a low abundance long non-coding RNA transcript, which is more comprehensively covered with use of KAPA Stranded RNA-Seq Kit with RiboErase (green) for library preparation. In comparison, Illumina Ribo-Zero Gold (orange) and NEBNext (blue) show less even coverage of this transcript. Data on file.
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Unsurpassed sequencing data quality

  • Detection of more genes and unique transcripts
  • Accurate and clear identification of splice sites and alternative gene splicing
  • Improved coverage enabling better detection of difficult and GC-rich transcripts
Hierarchically clustered heatmap showing the correlations of gene expression levels between different laboratories and RNA input amounts. All correlations were greater than 0.93, with technical replicates produced in the same lab exceeding 0.98. Generally, all libraries from a single lab, including those produced from different input amounts, clustered together. The data indicates that the KAPA Stranded RNA-Seq Kit with RiboErase delivers robust, reproducible, and unbiased gene expression data from a wide range of input amounts. Data on file.
0.973) which are better than libraries prepared from the equivalent kits from Illumina or NEB. Data on file."> Correlation of gene expression levels between replicate samples. Libraries prepared using 100 ng of UHR Total RNA with the KAPA Stranded RNA-Seq Kit with RiboErase show a very high correlation (R2>0.973) which are better than libraries prepared from the equivalent kits from Illumina or NEB. Data on file.
0.973) which are better than libraries prepared from the equivalent kits from Illumina or NEB. Data on file."> Correlation of gene expression levels between replicate samples. Libraries prepared using 100 ng of UHR Total RNA with the KAPA Stranded RNA-Seq Kit with RiboErase show a very high correlation (R2>0.973) which are better than libraries prepared from the equivalent kits from Illumina or NEB. Data on file.
0.973) which are better than libraries prepared from the equivalent kits from Illumina or NEB. Data on file."> Correlation of gene expression levels between replicate samples. Libraries prepared using 100 ng of UHR Total RNA with the KAPA Stranded RNA-Seq Kit with RiboErase show a very high correlation (R2>0.973) which are better than libraries prepared from the equivalent kits from Illumina or NEB. Data on file.
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Highly reproducible sequencing results

  • High correlation even between different testing conditions
  • Low sample-to-sample variation for more reliable results
Improved coverage of the whole transcript. The housekeeping gene Beta-Actin has improved coverage at the 3’ and 5’ regions using the KAPA Stranded RNA-Seq Kit with RiboErase (green) when compared to the equivalent Illumina Ribo-Zero Gold (orange) and NEBNext (blue) kits. Data on file.
Minimal positional bias. With 100 ng of high-quality RNA input, 3 positional coverage bias is minimized. Compared to Illumina Ribo-Zero Gold (orange) and the undepleted sample (magenta), KAPA Stranded RNA-Seq Kit with RiboErase (green) produces equivalent 5’ – 3’ coverage across transcript. When compared to NEBNext (blue), the KAPA Stranded RNA-Seq Kit with RiboErase provides more uniform distribution of reads per transcript. Data on file.
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Improved coverage uniformity

  • Uniform distribution of reads over each transcript
  • Minimal 5’ – 3’ bias across transcripts


*Data on file.

Related Products

Are you sequencing low-input, FFPE or high quality DNA?  RNA?  Check out these Kapa NGS products to improve your workflow and results:

KAPA Stranded mRNA-Seq Kits

KAPA Stranded mRNA-Seq Kits

KAPA RNA HyperPrep Kits with RiboErase

KAPA RNA HyperPrep Kits with RiboErase

KAPA mRNA HyperPrep Kits

KAPA mRNA HyperPrep Kits

Library Amplification

KAPA Library Amplification Kits

Library Preparation

KAPA Hyper Prep Kits

Library Quantification

KAPA Library Quantification Kits

Applications:

Applications
  • Whole Transcriptome
  • Gene Expression
  • Transcript Discovery and Annotation
  • Total RNA-Seq

Kit Specifications and Contents / Storage:

Kit Specifications and Contents / Storage

Kits can be stored for up to 10 months at -20ºC.

Kits contain KAPA Hybridization Buffer, KAPA Hybridization Oligos (Human/Mouse/Rat), KAPA Depletion Buffer, KAPA RNase H, KAPA DNase Buffer, KAPA Dnase, KAPA Fragment, Prime and Elute Buffer (2X), KAPA 1st Strand Synthesis Buffer, KAPA Script, KAPA 2nd Strand Marking Buffer, KAPA 2nd Strand Synthesis Enzyme Mix, KAPA A-Tailing Buffer (10X), KAPA A-Tailing Enzyme, KAPA Ligation Buffer (5X), KAPA DNA Ligase, KAPA PEG/NaCl SPRI Solution, KAPA Library Amplification Primer Mix (10X) and KAPA HiFiHotStartReadyMix (2X).

Components

RIBO-Component Chart

Specifications

Spec
Description
Compatible Platform
Illumina HiSeq®, NextSeq® and MiSeq®
Library Type
RNA
Starting Material
High-quality Total RNA
Input Amount
100 ng - 1 µg

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