Kapa biosystems/KAPA mRNA HyperPrep Kits/KK8702/12 adapters x 40 µl each

Description:

KAPA Single-Indexed Adapter Kit, Set B (30 µM)

KAPA mRNA HyperPrep Kits

Single-Day RNA

The KAPA mRNA HyperPrep Kits utilize novel chemistry that enables the combination of enzymatic steps and fewer reaction purifications, resulting in a truly streamlined solution for the preparation of high-quality mRNA-seq libraries. mRNA capture beads are used prior to library preparation, which enriches for mature mRNA over non-polyadenylated species, such as ribosomal, precursor, and noncoding RNAs.

The strand-specific workflow is flexible – supporting library construction from lower-input amounts and degraded samples. Kits contain all reagents required for mRNA capture and library preparation, with the exception of KAPA Adapters (available separately). Benefits include:

single-day library construction, inclusive of mRNA capture
reduced hands-on and overall time through fewer enzymatic and reaction cleanups
flexible input of 50 ng – 1 µg total RNA*
maintain over 99% strand specificity*
KAPA Pure Beads included for reaction purifications

KAPA RNA HyperPrep Kits

KAPA RNA HyperPrep Kits leverage the same cDNA synthesis and library preparation workflow improvements as outlined for the KAPA mRNA HyperPrep Kits. KAPA RNA Hyper Prep Kits include all enzymes and buffers required for cDNA library preparation, but do not contain mRNA capture reagents or KAPA Adapters. Kits can be used to prepare libraries from 1 ng – 100 ng of total, poly(A)-selected, or rRNA-depleted RNA.

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

The KAPA mRNA HyperPrep Kit results in a reduction in the total number of reads wasted due to both PCR duplicates and alignment to rRNA loci in comparison to the TruSeq and NEBNext kits. Libraries were generated in quadruplicate with 50 ng of high-quality Universal Human Reference (UHR) RNA using the manufacturers’ standard recommendations per workflow, where possible. Sequencing was performed using an Illumina HiSeq® 2500 in High Output mode with v4 chemistry and 2 x 100 bp read length. Reads aligning to rRNA were removed and paired reads were randomly subsampled to 14M for comparative analyses, including marked duplicates. Data on file.

With an equivalent amount of sequencing, more genes and unique transcripts are identified using the KAPA mRNA HyperPrep Kit in comparison to the TruSeq and NEBNext kits. Libraries were generated in quadruplicate with 50 ng of high-quality UHR RNA using the manufacturers’ standard recommendations per workflow, where possible. Sequencing was performed using an Illumina HiSeq® 2500 in High Output mode with v4 chemistry and 2 x 100 bp read length. Reads aligning to rRNA were removed and paired reads were randomly subsampled to 14M for comparative analyses, including marked duplicates. Data on file.

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Sequence what matters

Waste fewer reads due to the combination of rRNA carryover and PCR duplicates
Identify more unique transcripts and genes with equivalent sequencing

For the top 1000 transcripts, the KAPA mRNA Hyper Prep Kit results in more even coverage across transcript lengths, as assessed by both a normalized coverage plot and coverage coefficient of variation (CV), in comparison to competitor workflows. Libraries were generated with 50 ng of high-quality UHR RNA using the manufacturers’ standard recommendations per workflow, where possible. Sequencing was performed using an Illumina HiSeq® 2500 in High Output mode with v4 chemistry and 2 x 100 bp read length. Reads aligning to rRNA were removed and paired reads were randomly subsampled to 14M for comparative analyses, including marked duplicates. Data on file.

Improved coverage of GC-rich regions (outlined in red) of the YBX1 (rRNA depletion) and SLC2A4RG (mRNA capture) genes, along with improved splice junction and antisense transcript detection was demonstrated using the KAPA mRNA Hyper Prep Kit in comparison to competitor workflows. Libraries were generated with 50 ng of high-quality UHR RNA using the manufacturers’ standard recommendations per workflow, where possible. Sequencing was performed using an Illumina HiSeq® 2500 in High Output mode with v4 chemistry and 2 x 100 bp read length. Reads aligning to rRNA were removed and paired reads were randomly subsampled to 14M for comparative analyses, including marked duplicates. Data on file.

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Achieve superior coverage uniformity

Obtain more uniform distribution of reads across transcripts
Improve coverage of difficult GC-rich transcripts

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 RNA HyperPrep Kits with RiboErase

KAPA Stranded mRNA-Seq Kits

KAPA Library Amplification Kits

KAPA Hyper Prep Kits

KAPA Library Quantification Kits

Applications:

Applications

Gene expression analysis
Detection of gene fusions, isoforms, and other structural variants
Novel transcript identification, including noncoding transcripts
SNV discovery
Whole transcriptome (KAPA RNA Hyper Prep Kits only)
Targeted transcriptome (KAPA RNA Hyper Prep Kits only)

Kit Specifications and Contents / Storage:

Kit Specifications and Contents / Storage

Enzymes and buffers for rRNA depletion, cDNA synthesis, and library preparation can be stored for up to 10 months at -20°C. mRNA capture reagents and KAPA Pure Beads can be stored for up to 10 months at 4°C. (US only)

Kits include reagents for RNA fragmentation, cDNA synthesis, and library preparation. Kits with reagents for mRNA capture are also available.

RNAHyper_Component Chart

Specifications

Compatibility Platform

Illumina HiSeq, NextSeq, and MiSeq

Library Type

RNA

Starting Material

RNA HyperPrep Kit: High-quality total RNA, mRNA, or rRNA-depleted RNA; mRNA HyperPrep Kit: High-quality total RNA