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Pacbio RNA Sequencing
Nowadays, short-read RNA sequencing (RNA-seq) is the most widely used high-throughput method and has greatly expanded our ability to study transcriptome complexity, such as predicting alternative splicing events. However, short-read RNA-seq has many inherent limitations and cannot accurately determine all isoforms. In addition, this traditional approach does not handle well non-model organisms that lack well-annotated genomes in terms of structure and function. Long-read RNA-seq can overcome some of the inherent limitations of short-read RNA-seq and improve gene prediction and annotation in genome assembly, ultimately providing full-length mRNA sequences and complete structural information.
Based on PacBio's Single Molecule, Real-Time (SMRT) Sequencing technology, the average read length can now reach 80 Kb, which is already longer than the length of typical genes in the general transcriptome. Therefore, using this long-read sequencing platform for transcriptome research, full-length transcript information can be obtained directly without assembly, thus ensuring the accuracy of transcriptome sequencing results to a greater extent.
Advantages of Long-Read RNA-Seq over Short-Read RNA-Seq
- PacBio's SMRT Sequencing and Iso-seq analysis allow generating full-length transcripts with no assembly required.
- Long-read RNA-seq performs better in genomic non-model organisms that lack good annotation.
- Overcome the problem of short splicing and incomplete information of transcripts in species without a reference genome.
- It can realize the investigation of structural variants such as alternative splicing and fusion genes with reference genomic species.
Workflow of Our Pacbio RNA-Seq Service
Sample Requirements and Preparation
- RNA amount ≥ 5μg.
- RNA concentration ≥ 300ng/μL, OD260/280 =1.8 ~2.0.
- All RNA samples are validated for purity and quantity.
Data Analysis Contents
As one of the leading international providers of long-read RNA-seq services, we provide comprehensive, customized experimental protocols and professional individualized analytical protocols for specific species, research objectives, and budgets. Our specific analysis services include,
| Items (without reference genome) | Items (with reference genome) |
| -Data QC -Transcript classification -Transcript clustering and correction -Functional annotation of full-length transcript sequences -LncRNA analysis -CDS prediction |
-Data QC -Transcript classification -Transcript clustering and correction -Reference genome alignment -Alternative splicing analysis -Novel RNA prediction -New transcript isoform annotation -Fusion gene analysis -LncRNA analysis -CDS prediction |
Benefits of Our Services
- Advanced experimental platform. Stable running sequencing platform that produces high-quality sequencing data and provides faster project cycle time.
- Professional Experimental consultation and design. Senior technical team to guarantee high-quality research results.
- Years of experience in long-read RNA seq analysis. International long-read sequencing pioneer. We have provided accurate and affordable long-read RNA-seq services for years.
Related Services
- Full-Length Transcript Sequencing (Iso-Seq)
- Animal/Plant RNA Sequencing
- Long-Read Sequencing of RNA Methylation
- Full-Length Transcriptome Profiling Combined with Long- and Short-Read Sequencing
- Full-Length Spatial-Temporal Transcriptome Profiling
- Single-Cell Full-Length Transcriptome Sequencing
CD Genomics is capable of providing long-read RNA sequencing service packages using PacBio Iso-seq analysis and Oxford nanopore sequencing. Our service pipeline can be tailored to your research interest. Please don’t hesitate to contact us. We look forward to working with you on your next project.