Whole transcriptome sequencing

Overview

• Library prep kits designed for use with the xGen mRNA Enrichment Module and xGen Hyb Capture
• Efficiently conduct expression profiling and fusion detection experiments thanks to high mapping rates, low duplicate rates, high number of genes detected, and comprehensive transcript coverage
• Accommodates your samples with a single kit for consistent results across input range. Save time, reduce costs, and produce consistent libraries
• Adaptase technology eliminates need for adapter titration resulting in minimal adapter dimers and high strandedness.  Go from RNA to library in as little as 3.5 hours!
• Further reduce costs by pooling with revolutionary normalization technology – xGen Normalase reduces sequencing costs, saves time and increases throughput with uniform sampling and fewer handling steps to generate balanced library representation allowing higher multiplexing per run and obtain predictable read numbers.

What is whole transcriptome sequencing?

Whole transcriptome sequencing is an RNA-seq approach used to profile RNA transcripts across a sample. It helps researchers measure gene expression, compare biological conditions, and study transcript-level changes such as alternative splicing, isoform usage, and gene fusions.

Because total RNA is dominated by ribosomal RNA, which can make up roughly 80–98% of RNA molecules in a sample, enrichment or depletion is often used before library preparation to focus sequencing reads on more informative transcript content. In an IDT workflow, the xGen mRNA Enrichment Module enriches poly(A)-tailed transcripts, helping economize sequencing by focusing on mature, coding mRNAs for mRNA-seq workflows. The workflow is also compatible with upstream ribodepletion approaches when study goals or sample quality require retention of non-polyadenylated transcripts.

RNA is then converted into sequencing-ready, stranded libraries using the xGen RNA Library Prep Kit or xGen Broad-Range RNA Library Prep Kit. Both kits use Adaptase™ technology, which adds a single-stranded R2 Stubby Adapter to the 3’ end of first-strand cDNA and eliminates the need for second-strand cDNA synthesis and degradation. The xGen RNA Library Prep Kit supports a faster workflow with fewer steps for 100 ng – 1 µg total RNA, while the xGen Broad-Range RNA Library Prep Kit supports broader input flexibility from 10 ng – 1 µg total RNA.

For high-throughput labs, 96-reaction product configurations include the xGen Normalase Module and xGen Normalase UDI Primer Pairs to support enzymatic library normalization and multiplexing. This approach reduces the need for individual sample quantification and manual equimolar pooling, generating normalized library pools ready for Illumina-based sequencing. The workflow can also be combined with compatible xGen hybridization capture reagents for targeted mRNA capture sequencing studies.

Method Data

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