Stranded mRNA-Seq Kits

KAPA Stranded mRNA-Seq Kits

For Illumina Sequencers

Available internationally on April 11, 2014

Even difficult messages should be understood.

The KAPA Stranded mRNA-Seq Kit generates libraries from 100 ng – 4 µg of total RNA with greater than 99% strand specificity and superior sequence quality. Kits are optimized for the improved coverage of GC-rich and low-abundance transcripts, resulting in the identification of more genes. Kits contain KAPA HiFi for high-efficiency and low bias library amplification, as well as KAPA mRNA Capture Beads and a streamlined, "with-bead" protocol.

Applications:

• Gene expression
• Single nucleotide variation (SNV) discovery
• Post-transcriptional SNVs
• Fusion gene identification
• Targeted transcriptome
• Whole transcriptome

Figure 1: Improved coverage of GC-rich transcripts.
The 5' and 3' exons (outlined in red) of the DVL3 transcript contain regions of very high GC content. These regions are covered to a significantly greater depth by the KAPA Stranded mRNA-Seq Kit in comparison to the Illumina® TruSeq™ Stranded mRNA Sample Prep Kit.

Figure 2: Improved coverage of low-abundance transcripts
GLTPD1, a lesser-expressed transcript, is covered more comprehensively with the KAPA Stranded mRNA-Seq Kit at 500 ng input total RNA (outlined in red). In comparison, Illumina TruSeq Stranded mRNA Sample Prep Kit shows coverage gaps, even with higher inputs (4 μg).

Figure 3: Highly mapped reads and strand-specificity enable sensitive detection of expressed genes
With similar numbers of filter-passed reads (>30M), KAPA Stranded mRNA-Seq libraries generate a greater percentage of mapped reads and lower duplication rates than analogous libraries prepared with the Illumina Stranded mRNA-Seq Sample Prep Kit while maintaining 99% strand-specificity.

Figure 4: Minimal positional bias
With intact, high-quality RNA input, 3' positional coverage bias was minimized (average across top 1000 transcripts shown). Compared to the Illumina TruSeq Stranded mRNA Sample Prep Kit, KAPA Stranded mRNA-Seq Kit produces equivalent 5’ – 3’ coverage across transcript.

Code	Description	Kit Contents
Stranded RNA-Seq Kits - without bead
KK8400	Kapa Stranded RNA-Seq Kit - 24 rxn	Includes buffers and enzymes required for construction of stranded RNA-Seq libraries via the following steps: fragmentation, 1st and 2nd strand cDNA synthesis, and cDNA library construction, including A-tailing, adapter ligation, and library amplification.
KK8401	Kapa Stranded RNA-Seq Kit - 96 rxn	Contains all of the buffers and enzymes required for construction of stranded RNA-Seq libraries via the following steps: fragmentation, 1st and 2nd strand cDNA synthesis, and cDNA library construction, including A-tailing, adapter ligation, and library amplification.
Stranded mRNA-Seq Kits - mRNA Capture Beads
KK8420	KAPA Stranded mRNA-Seq Kit - 24 rxn	Contains all of the buffers and enzymes required for construction of stranded RNA-Seq libraries via the following steps: mRNA capture, fragmentation, 1st and 2nd strand cDNA synthesis, and cDNA library construction, including A-tailing, adapter ligation, and library amplification.
KK8421	KAPA Stranded mRNA-Seq Kit - 96 rxn	Contains all of the buffers and enzymes required for construction of stranded RNA-Seq libraries via the following steps: mRNA capture, fragmentation, 1st and 2nd strand cDNA synthesis, and cDNA library construction, including A-tailing, adapter ligation, and library amplification.