- Fast and reliable—get RNA-seq libraries ready for hybridization capture in 3.5 hours
- Provides comprehensive data—for mapping, on-target, and transcript coverage
- Compatible with a range of input types and quantities—prepare RNA-seq libraries from low-input and/or low-quality RNA samples using the xGen Broad‑Range RNA Library Prep Kit
- Consistent libraries—minimal adapter dimers, so adapter titration is not required
- Variety of indexing options available—for a total of 1536 Unique Dual Index (UDI) primer pairs
- Flexible and customizable—choose between different hybridization panels depending on your research needs
- Automation-friendly RNA-seq workflow
What is targeted RNA sequencing?
Targeted RNA sequencing (RNA-seq) is a cost-effective tool for deep sequencing of specified regions of interest within the transcriptome. This approach provides deep sequencing of targeted regions while omitting the undesired regions that often result
in a high number of sequencing reads not relevant to the research study.
Reliable, time- and cost-effective approach to targeted RNA-seq workflow
RNA-seq provides the researcher with data about the RNA molecules in the sample at a specific time point—a snapshot in time. In short, the workflow consists of RNA extraction, rRNA depletion or mRNA enrichment, cDNA conversion, library preparation,
and finally, sequencing. Sometimes identifying all RNAs in a sample is beyond the scope of the experiment, and a strategy to focus the library to regions or targets of interest is preferred. In targeted RNA-seq, the traditional workflow can be modified
to consist of RNA extraction, cDNA conversion, and library preparation, followed by hybridization capture with either a custom or predesigned hyb cap panel.
For library prep, IDT offers two different solutions that include cDNA conversion.
- For regular RNA samples, the IDT xGen RNA Library Prep Kit offers a fast NGS transcriptomic research workflow. RNA-seq libraries can be produced using tissue samples, blood, or high-quality RNA samples with an RNA Integrity Number (RIN) of 7–10. This RNA-seq workflow also utilizes Adaptase™
technology to produce libraries following first-strand cDNA synthesis (see the xGen RNA Library workflow).
This workflow can be easily combined with xGen UDI Primer Plates for
- The xGen Broad‑Range RNA Library Prep Kit supports a wide input range, including low-quality formalin-fixed paraffin-embedded (FFPE) RNA samples. This stranded RNA-seq workflow for low-quality and/or low-input RNA samples utilizes Adaptase technology to produce libraries following first-strand
cDNA synthesis (refer to xGen Broad-Range RNA Library workflow).
Samples are recommended to have a RIN >2 or DV200 >30 (DV200 is a quality metric that represents the percentage of RNA fragments longer than 200 nucleotides). This workflow can also be easily combined with xGen UDI Primer Plates for indexing.
After library prep, hybridization capture with IDT xGen Hyb Probes, which are individually synthesized, 5’-biotinylated oligos, enriches for fragments corresponding to the targets of interest. For researchers interested in the mRNA subset expressed
from exon regions of the genome, you can pair the xGen Broad‑Range RNA Library Prep Kit or the xGen RNA Library Prep Kit with the xGen Exome Hyb Panel v2 for more efficient expression profiling, yielding a higher percentage of coding bases and a lower percentage of intronic bases than whole transcriptome data (see Figure 1B).
For researchers interested in a more focused target space, IDT offers a variety of other xGen Predesigned Hybridization Capture Panels,
and if these are not exactly what you want, an IDT xGen Custom Hyb Panel can be designed to target your specific genetic regions of interest. IDT also offers the xGen Hybridization and Wash v2 Kit that includes buffers, Cot DNA, our xGen 2X HiFi PCR Mix, and streptavidin-coated magnetic beads (note, the beads and reagents cannot be ordered separately).
In order to prevent non-specific pull-down of fragments during the hybridization capture reaction, IDT has developed xGen Universal Blockers that inhibit adapter:adapter annealing that can be one of the causes of non-specific interactions.