iGeneTech RNA Exome is a capture product targeting the human whole transcriptome. Designed based on the RefGene and RefSeq databases, it enables targeted capture of a 46.1 Mb region, including coding regions of 69,209 transcripts from 20,761 genes, as well as partial UTR regions. Compared to whole transcriptome sequencing, it achieves more efficient enrichment of target regions.
It can also efficiently enrich small amounts of degraded RNA (such as FFPE samples) and low-expression, rare transcripts, thereby obtaining more valid information. In clinical testing and translational research, it facilitates studies on RNA-level fusion events and differential expression, providing reliable transcriptome sequencing data support for disease research.
Design Protocol
Compared with the DNA-level probe design scheme, iGeneTech's RNA-level design uses transcripts as the reference sequence. It fully aligns with the sequence characteristics of RNA samples, eliminates the influence of sequences in intron regions, and additionally incorporates exon boundary probes. This fully ensures the detection of transcripts with different splice sites and fusion events.
Product Features
Unlike DNA-level design and research, RNA Exome is meticulously designed in terms of reference sequences, design schemes, and region selection.
Comprehensive coverage: Full coverage of coding regions of 69,209 transcripts from 20,761 genes.
Targeted enhancement: Probes on both edges of exons supplement to improve fusion detection capability.
Precise targeting: Based on fusion-related information in Fusion GDB2, relevant UTR regions are prioritized for coverage.
TCR/BCR: Additional TCR/BCR probes are included.
Probe filtering: Probe design filters out rRNA and globin-related genes.
Test Data
01 Efficient Capture of Target Regions
Figure 1. Capture Performance Test of RNA StandardsUsing Shuimu Jiheng RNA standards, libraries were constructed with an RNA library construction kit at input amounts of 10 ng and 200 ng. After hybrid capture, sequencing was performed on an Illumina NovaSeq 6000 with PE150 (paired-end 150) reads.
02 Significant Results in CDS Region Enrichment Analysis
Figure 2. Proportion of Sequencing Data Aligned to CDS Regions Across Different Technical ProtocolsWith 10 ng of total RNA input, detection was performed using RNA Seq and RNA Exome respectively, and the sequencing data was aligned to CDS region annotations.
03 Sensitive Detection of Fusion Genes
Table 1. Detected Fusion Results of Positive Fusion Standards
Note: Results of the comparative analysis on the detection of fusion events in positive standards showed that, under the condition of using the same number of positive standards but different input amounts, all fusion events were detected, which was consistent with the expected results.
Product Info
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