RNA Sequencing
/in NGS, Partners, Twist/by Harshita SharmaPRODUCTS
RNA Sequencing
Twist’s RNA sequencing workflows offer a complete NGS solution that produces uniform libraries for RNA sequencing.
Twist offers targeted and whole transcriptome workflows that reduce time at the bench and integrate with our current set of NGS reagents, including our target enrichment kits. Each workflow delivers high quality libraries ready for sequencing from a wide range of inputs and sample types, including translation research samples.
Twist offers two comprehensive RNA sequencing workflows: Targeted RNA Sequencing and Whole Transcriptome Sequencing. The Targeted RNA Sequencing workflow involves creating custom panels to focus on specific RNA transcripts, utilizing streamlined library preparation kits, unique dual indices, molecular identifiers, optimized target enrichment, and a proprietary exon-aware panel design algorithm. This allows for sensitive and efficient sequencing, even with low-quality RNA from FFPE samples.
On the other hand, the Whole Transcriptome Sequencing workflow measures expression levels of various RNA species, including mRNA and lncRNA. It involves preparing libraries from total RNA extracted from both fresh and FFPE samples using Twist RNA Library Prep and Twist rRNA & Globin Depletion kit. This workflow enables RNA Library Preparation in less than 5 hours, providing a comprehensive view of the entire transcriptome. Two charts accompany the descriptions, detailing the targeted and whole transcriptome sequencing workflows.
The collaboration of Twist RNA Exome, Twist RNA Library Prep, and Twist Target Enrichment forms a reliable toolkit for transcriptome sequencing, accommodating RNA from diverse sources, including FFPE samples. Notably, the RNA Exome enhances signal strength while requiring fewer sequencing reads, enabling the detection of low-expressing targets crucial for accurate transcriptional profiling. Its exon-aware design approach allows the identification of isoforms and junctions often overlooked in conventional methods, delivering precise and uniform sequencing reads tailored for analyzing protein-coding sequences within the human transcriptome.
Designed exclusively for the human transcriptome, the RNA Exome targets 35.8 Mb bases, 19,708 genes, and 63,215 isoforms using sequences from Gencode and RefSeq. With over a 1.8-fold enrichment compared to whole transcriptome sequencing, it features a target enrichment approach built for RNA, incorporating an exon-aware probe design for protein coding regions, fusions, and isoforms. This design not only ensures precision but also reduces reads per sample, facilitating efficient analysis of multiple samples. The RNA Exome is versatile, compatible with FFPE samples and low RNA input. In practical tests, it demonstrates the capability to detect more targets with fewer reads across a range of RNA inputs, including FFPE and universal human reference RNA, using libraries prepared with the Twist RNA Library Prep.
The Twist Alliance CeGaT RNA Fusion Panel Kit emerges as a crucial tool in the precise detection of gene fusions, particularly impactful oncogenic driver mutations in various cancer types. Rapid and accurate detection holds significant clinical implications, guiding treatment decisions effectively. This RNA-based enrichment panel stands out by offering an enhanced targeted and sensitive approach, surpassing traditional methods.
Curated in collaboration with CeGaT, a renowned genetic diagnostic and sequencing company in Europe, this panel encompasses 160 fusion genes associated with approximately 30 cancer types. Uniquely, it is designed not only for known gene fusions but also to unveil novel ones. The panel’s efficiency is amplified when used in conjunction with the Twist RNA Library Prep Kit, ensuring an end-to-end workflow with exceptional performance.
With an optimized design featuring 7394 probes and a breakpoint design for 66 genes, the panel excels in covering a diverse array of cancer types. The incorporation of Twist core enrichment technology further maximizes capture efficiency, instilling confidence in RNA gene fusion detection. This panel is particularly well-suited for screening oncology samples, providing critical insights for treatment decisions or uncovering novel fusions for tumor classifications.
*Note: The Twist Alliance CeGaT RNA Fusion Panel – 3 MB holds ISO-13485 certification, reinforcing its reliability in clinical applications.
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