Sample Submission Guidelines
What is polyA Sequencing?
Poly(A) sequencing provides a powerful window into the life cycle of mRNA molecules by precisely measuring the length and composition of poly(A) tails located at the 3' end of transcripts. These tails play crucial roles in:
- mRNA Stability
Longer poly(A) tails often protect mRNA from degradation, extending transcript lifespan in the cell. - Translational Efficiency
Poly(A) length influences how effectively mRNA is translated into proteins. - Gene Expression Regulation
Shifts in poly(A) tail length reflect dynamic biological processes, such as development, stress responses, and disease states.
Traditional methods like LC-MS or gel electrophoresis provide only average poly(A) length estimates. In contrast, sequencing-based technologies—including TAIL-seq and direct RNA sequencing—deliver base-level resolution and detect transcript-specific variations and 3' end modifications. This level of detail empowers researchers to uncover fine-scale regulatory mechanisms and support applications such as mRNA therapeutic development and biomarker discovery.
CD Genomics polyA Sequencing Services
At CD Genomics, we offer comprehensive polyA sequencing services designed to deliver high-resolution insights into mRNA biology. Utilizing advanced platforms—including TAIL-seq and direct RNA sequencing—we empower researchers to precisely measure poly(A) tail length and identify 3'-end modifications across the transcriptome.
Our polyA sequencing services help you:
✅ Quantify poly(A) Tail Lengths with Nucleotide Precision
Obtain accurate measurements for each transcript, avoiding biases introduced by oligo(dT) enrichment or PCR amplification.
✅ Detect 3'-End Modifications and Variants
Reveal uridylation, guanylation, and other non-adenine additions that influence mRNA decay or translational efficiency.
✅ Profile Alternative Polyadenylation Sites
Map poly(A) sites genome-wide to understand transcript isoform diversity and regulatory dynamics.
✅ Support mRNA Therapeutic Development and QC
Ensure poly(A) tail integrity and length uniformity for in vitro transcribed (IVT) mRNA products, critical for therapeutic efficacy and regulatory compliance.
✅ Uncover Regulatory Mechanisms in Health and Disease
Explore poly(A) tail dynamics in diverse biological contexts, from embryonic development to cancer and plant biology.
Workflow
Service Specifications
| Feature | Details |
|---|---|
| Sample Type | Total RNA (≥1–2 μg), high integrity (RIN >7 recommended) |
| Input Quality | DNase-treated, free from inhibitors and contaminants |
| Poly(A) Tail Length Range | Detectable from ~10 nucleotides to >250 nucleotides |
| Sequencing Platforms | Illumina (TAIL-seq), Nanopore Direct RNA Sequencing |
| Detection Capabilities | Poly(A) tail length, alternative polyadenylation sites, 3'-end modifications |
| Data Output | FASTQ files, poly(A) tail length reports, visualization plots |
| Bioinformatics Analysis | Tail length quantification, differential analysis, isoform-specific insights |
| Applications | mRNA stability studies, translational efficiency, mRNA therapeutics QC, alternative polyadenylation profiling, disease research |
| Service Type | Research Use Only (RUO) |
Bioinfomatics Analysis
Technical Highlights
CD Genomics combines cutting-edge sequencing technologies with rigorous bioinformatics to deliver comprehensive polyA tail analysis. Our polyA sequencing platform offers unique technical strengths:
High-Resolution Tail Length Measurement
- Achieve single-nucleotide resolution of poly(A) tail lengths for individual transcripts.
- Distinguish subtle differences in tail length that correlate with mRNA stability and translation efficiency.
Detection of 3'-End Modifications
- Identify uridylation, guanylation, and mixed non-adenine additions at mRNA 3' ends.
- Reveal novel regulatory signals affecting mRNA degradation or translational repression.
No Oligo(dT) Bias
- Avoid biases introduced by oligo(dT) enrichment, enabling accurate detection of short poly(A) tails.
- Capture full-length poly(A) tails without sequence artifacts.
Transcript-Level Resolution
- Map poly(A) tails directly to specific transcripts.
- Analyze isoform-specific poly(A) dynamics critical for alternative polyadenylation studies.
Advanced Sequencing Platforms
TAIL-seq:
- Paired-end sequencing approach correlating 3' tail length with gene identity.
- Ideal for studies requiring precise poly(A) tail length profiling and modification detection.
Direct RNA Sequencing:
- Nanopore-based technology reads native RNA molecules.
- Eliminates PCR and reverse transcription biases.
- Simultaneously profiles poly(A) length and RNA modifications.
Robust Bioinformatics Pipeline
- Automated detection and quantification of poly(A) tail lengths.
- Differential analysis between experimental conditions.
- Visualization outputs such as histograms, boxplots, and scatter plots for intuitive data interpretation.
Applications of polyA Sequencing
mRNA Stability and Decay Studies
- Measure poly(A) tail lengths to infer transcript half-lives and degradation kinetics.
- Identify transcripts undergoing rapid turnover or exhibiting stable expression profiles.
Translational Efficiency Analysis
- Correlate poly(A) tail length with ribosome loading and protein production rates.
- Uncover mechanisms that fine-tune gene expression at the post-transcriptional level.
Alternative Polyadenylation Profiling
- Map poly(A) sites genome-wide to study transcript isoform diversity.
- Investigate how shifts in polyadenylation affect gene regulation in development, stress response, and disease
mRNA Therapeutic Development and Quality Control
- Evaluate poly(A) tail length distribution in in vitro transcribed (IVT) mRNA products.
- Ensure compliance with regulatory expectations for mRNA-based therapeutics and vaccines.
Disease Research and Biomarker Discovery
- Explore changes in poly(A) tail dynamics associated with cancer, neurodegeneration, and other pathologies.
- Identify transcript-specific poly(A) signatures as potential diagnostic or prognostic biomarkers.
Plant and Agricultural Genomics
- Analyze tissue-specific poly(A) tail patterns in crops and model plants.
- Investigate poly(A) tail dynamics in stress responses, growth, and developmental processes.
By enabling precise, transcript-level analysis of poly(A) tails, our services help researchers decode key regulatory layers of gene expression and advance discoveries across multiple scientific fields.
Deliverables
✅ Raw Sequencing Data
- FASTQ files containing high-quality reads for downstream analysis.
- Compatible with standard bioinformatics pipelines.
✅ Poly(A) Tail Length Profiles
- Detailed reports quantifying poly(A) tail lengths across transcripts.
- Distribution plots to visualize tail length variability and detect global shifts.
✅ Alternative Polyadenylation Insights
- Identification of multiple poly(A) sites within genes.
- Isoform-specific poly(A) tail length analysis for alternative polyadenylation studies.
✅ 3'-End Modification Detection
- Reports highlighting non-adenine additions (e.g., uridylation, guanylation) at transcript 3' ends.
- Insights into regulatory mechanisms affecting mRNA stability and translation.
✅ Differential Analysis Between Conditions
- Statistical comparison of poly(A) tail length distributions across experimental groups.
- Identification of significant changes linked to biological processes or treatments.
✅ Data Visualization Outputs
Publication-ready plots, including:
- Histograms
- Boxplots
- Scatter plots
- Visual tools to support interpretation and presentation of poly(A) data.
✅ Expert Bioinformatics Support
- Assistance with data interpretation and custom analyses.
- Tailored recommendations for follow-up experiments.
Sample Requirements
| Sample Type | TAIL Iso-seq Requirement |
|---|---|
| Total RNA | ≥ 1 μg |
| Fresh animal tissue (dry weight) | ≥ 300 mg |
| Fresh plant tissue (dry weight) | ≥ 500 mg |
| Fresh cultured cells | ≥ 1 × 10⁷ cells |
| Fresh whole blood | 4–5 ml |
| Microorganisms | ≥ 1 × 10⁷ cells or ≥ 500 mg |
FAQs
1. What types of samples can I use for polyA sequencing?
Any high-quality total RNA (RIN > 7, ≥1–2 µg) across species—including animals, plants, and microbes—is suitable. No poly(A) enrichment is required.
2. Can you accurately measure short poly(A) tails?
Yes. Unlike oligo(dT)-based methods, our polyA sequencing platform reliably quantifies tails as short as ~10 nucleotides.
3. What does APA (Alternative Polyadenylation, PAA) analysis tell me?
APA reveals different poly(A) site usage within a gene—key for understanding transcript isoform diversity, 3' UTR length regulation, and its impact on mRNA stability or protein production.
4. Can we identify 3'-end modifications like uridylation or guanylation?
Absolutely. Our method detects non‑adenine additions at the 3' end, offering insights into mRNA regulation that LC‑MS or gel methods cannot provide.
5. Is bioinformatics analysis of polyA data included?
Yes. We provide both standard and customized reports: tail length distribution, APA site mapping, differential poly(A) analysis, and expert consultation for interpretation.
6. Are these services suitable for clinical diagnostics?
No. Our polyA sequencing service is for Research Use Only (RUO) and not intended for clinical or diagnostic applications.
Title: Nano3P-seq Unlocks mRNA Expression Insights and Poly(A) Tail Dynamics via End-Capture Nanopore Sequencing
Polyadenylation—the addition of poly(A) tails—is a crucial process shaping RNA maturation, lifespan, and stability. As organisms develop, these tails change in length, directly influencing how efficiently mRNA is translated into proteins.
In a study published in Nature Methods (Impact Factor 36.1) in January 2023, researchers introduced Nano3P-seq, a cutting-edge technique leveraging Nanopore's sequencing platform. This end-capture approach not only quantifies RNA abundance across the transcriptome but also dissects poly(A) tail composition and length dynamics—all without requiring PCR amplification or RNA adapter ligation.
Unlike traditional sequencing methods, Nano3P-seq uses a template-switching mechanism to read RNA molecules from their 3' ends. This design enables precise profiling regardless of whether transcripts carry poly(A) tails. The team demonstrated that Nano3P-seq effectively estimates RNA levels and tail lengths while capturing diverse RNA species, including mRNA and long non-coding RNAs.
Strikingly, the study revealed that poly(A) tails appear even in unexpected regions, such as the 16S mitochondrial ribosomal RNA in mouse and zebrafish models. Furthermore, the data highlighted dynamic regulation of poly(A) tail length during vertebrate embryonic development—a factor intimately tied to mRNA degradation and stability.
One standout capability of Nano3P-seq is its ability to detect non-adenine bases embedded within poly(A) tails in single sequencing reads. These non-adenine insertions, observed across various developmental stages, shed light on new regulatory layers influencing gene expression in vertebrate embryos.
Researchers and drug developers interested in RNA biology now have a powerful tool to explore not just transcript abundance but also subtle tail modifications that may hold clues for disease mechanisms or therapeutic targets.
References
- Begik O, Diensthuber G, Liu H, et al. Nano3P-seq: transcriptome-wide analysis of gene expression and tail dynamics using end-capture nanopore cDNA sequencing. Nature Methods, 2023.
- Jia J, Lu W, Liu B, et al. An atlas of plant full-length RNA reveals tissue-specific and monocots–dicots conserved regulation of poly (A) tail length. Nature Plants, 2022.
