Transcriptomics
We offer comprehensive RNA sequencing services, including total RNA, mRNA, and small RNA library preparation and sequencing on Illumina platforms. Our services are tailored to meet the specific needs of each project, and we also provide consultation services for protocol development and collaborative research.
Consultation Services:
We offer consultations for project guidance and collaboration on protocol development. Contact us to discuss your needs.
Important RNA Sample Handling Guidelines
To ensure the integrity of your RNA samples, limit the number of times you freeze and thaw them. Always keep RNA samples on ice during handling to prevent degradation. Please be aware that we cannot guarantee successful library preparation if the samples do not meet our specified requirements.
Measure RNA integrity using capillary electrophoresis methods (e.g., Fragment Analyzer, Bioanalyzer, or TapeStation) and provide the RIN values for each sample when submitting a project request. This analysis can also be performed at CTG.
Sample Delivery Instructions (for all services):
- Aliquot samples on dry ice in a fully skirted 0.2 ml plate with a cold-resistant seal.
- Place samples column-wise (A1, B1, C1...) and upload the sample sheet to iLab.
Our standard workflow uses the Illumina Stranded mRNA Prep, Ligation, which purifies and captures mRNA molecules from total RNA using oligo(dT) magnetic beads. This ensures strand orientation for detecting antisense transcription and enhances transcript annotation. The kit’s ligation-based method adds adapters and indexes to create high-quality, dual-indexed libraries compatible with all Illumina sequencing platforms. Sequencing is performed on NovaSeq X Plus, 2 x 150 bp read structure, targeting 25M reads per sample.
Sample Requirements for mRNA Profiling:
- Sample Type: Total RNA
- Concentration: 10 ng/μL. Quantified via fluorometric methods (e.g., Qubit). Nanodrop measurements are not accepted.
- Sample Quality: gDNA-free (DNase I treatment recommended). Resuspended in nuclease-free water.
- RIN Value: RIN > 8.0 recommended
- Volume: 35 μL
For more information, please contact CTG services or open a consultation project on iLab.
For whole-transcriptome analysis, we use the Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero kit. This solution depletes ribosomal RNA and globin RNA, enabling efficient transcriptome sequencing even from low-input and low-quality samples. The kit supports both coding and non-coding RNA detection. Sequencing is performed on NovaSeq X Plus, 2 x 150 bp read structure, targeting 50M reads per sample.
Sample Requirements for Total RNA Sequencing:
- Sample Type: Total RNA
- Concentration: 20 ng/μL. Quantified via fluorometric methods (e.g., Qubit). Nanodrop measurements are not accepted.
- Sample Quality: gDNA-free (DNase I treatment recommended). Resuspended in nuclease-free water.
- RIN Value: RIN > 8.0 recommended
- Volume: 15 μL
For more information, please contact CTG services or open a consultation project on iLab.
For low input or degraded samples, we offer the TaKaRa SMARTer Stranded Total RNA-Seq Kit v3, optimized for RNA inputs as low as 250 pg–10 ng or a few cells (10–1,000). This kit is ideal for FFPE samples and includes rRNA depletion and the integration of unique molecular identifiers (UMIs) to improve data accuracy. Sequencing is performed on NovaSeq X Plus, with read structure of 2 x 150 bp and aiming for 50M reads per sample
Sample Requirements for Pico Input RNA Sequencing:
- Sample Type: Total RNA
- Concentration: > 1 ng/μL
- Sample Quality: High quality or degraded RNA accepted. gDNA-free (DNase I treatment recommended). Resuspended in nuclease-free water.
- RIN Value: RIN > 7.0 recommended
- Volume: 12 μL (min)
For more information, please contact CTG services or open a consultation project on iLab.
We offer small RNA sequencing using the QIAseq miRNA UDI Library Kit for precise library preparation. This kit uses unique dual indexes (UDIs) and incorporates unique molecular identifiers (UMIs) to improve data accuracy. It is Ideal for profiling miRNAs from a wide range of sample types, including low-input and degraded RNA. Sequencing is performed on NovaSeq X Plus, with read structure of 2 x 150 bp and aiming for 5M reads per sample
Sample Requirements for Small RNA Sequencing:
- Sample Type: small RNA
- Concentration: 20 ng/μL
- Sample Quality: gDNA-free (DNase I treatment recommended). Resuspended in nuclease-free water. Ensure extraction protocol retains small RNAs (<200bp)
- Volume Cells/Tissues: 10 µl
- Volume Serum/Plasma: 5 μL of RNA eluate (200 μL serum/plasma processed) using the miRNeasy Serum/Plasma Kit
For more information, please contact CTG services or open a consultation project on iLab.