Product introduction

1. Product application

● Gene expression analysis of high and low quality RNA samples (e.g. extracted from FFPE tissue).

● Discovery of single nucleotide variation (SNV).

● Recognition of splicing sites and fusion genes.

● Characterization of RNA without poly tails, including non-coding RNA and prerequisite RNA.

2 Product Advantages

● Up to 99.98% rRNA removal efficiency ● The initial total RNA range is 100ng to 1μg

● Suitable species: human, mouse, rat ● Reliable and reproducible results can be obtained at different starting amounts

● Work flow is easy to achieve automation, there are a variety of automatic platform docking schemes.

3. Advantage performance

1) Industry-leading rRNA removal. It can effectively remove rRNA from FFPE samples and reduce rRNA reads, which is more economical for sequencing and deeper for transcripts

2) Maximum coverage uniformity

● The distribution of all transcripts is uniform

●5 '-3' is less favorable

3) Unmatched sequencing data quality

● Detect more genes and specific transcripts

● Accurate and clear identification of splicing sites and selective splicing genes

● High coverage, better detection of difficult or high GC content transcripts

　High-quality sequencing data obtained by using the Kapa RNA-Seq Kit with Riboerase workflow, compared with the results of similar kits of Illumina and NEB;

The workflow of the Kapa Stranded RNA-seq Kit with Riboerase not only achieved the same sequencing read mapping rate, but also had low duplication rates and high coverage homogeneity, as well as detected more specific transcription segments and genes. Thus, the sensitivity of detecting expressed genes in samples is improved.

The coverage of GC enriched transcriptional fragments was improved:

The figure above shows the sequencing coverage and cleavage site recognition of haemoglobin alpha transcript(HBA1). The GC content of HbA1 is 62.8%. The areas with high GC content are marked red in the track showing GC content in the figure. The areas with high AT content were marked in blue, and the samples processed with a Kapa Stranded RNA-seq Kit with Riboerase workflow (green) had more uniform coverage than those processed with a similar Illumina Kit (orange) or NEB Kit (blue). More sensitive splice site recognition and a more representative data pattern compared to the untreated rRNA sample (red).

　The detection rate of low abundance transcriptional fragments is improved:

MALAT1, a non-coding RNA fragment with low abundance, could be uniformly covered by the Kapa RNA-seq Kit with RiboErase(green) in the library preparation process. In contrast, The Illumina-Zerogold (orange) and NebNext (blue) workflows were used to obtain a less uniform coverage of this transcription fragment.

4) Highly repeatable sequencing results

● Heat has a high correlation even under different test conditions

● The differences between samples are small and the results are reliable

　　Correlation of gene expression levels between repeated samples: All of the above experiments used 100ng and UHR total RNA as starting samples. When database building was done with the Kapa RNA-Seq Kit with Riboerase workflow, the gene expression water samples showed a very high correlation (R2>0.973) between duplicate samples than with Illumina or NEB kits of the same kind.

Related product information