RNA-Seq
SeqInsight is providing accurate and affordable RNA-Seq (RNA sequencing) services. We combine both Illumina (short reads) and PacBio (long reads) platforms to obtain the transcriptome that allows de novo assembly or re-sequencing for bacteria, plants, and animals.
The Introduction of RNA-Seq
RNA-Seq is the premier tool for mapping and quantifying transcriptomes by utilizing next-generation sequencing (NGS) technology. The transcriptome refers to the complete set of transcripts in a cell, which provides information on the transcript level for a specific developmental stage or physiological condition. Understanding the transcriptome is necessary for interpreting the functional elements of the genome and understanding development and disease. The key purpose of transcriptomics includes cataloging all species of transcripts; determining the transcriptional structure of genes; and quantifying the expression levels of each transcript under different conditions.
RNA-Seq delivers an unbiased and unprecedented high-resolution view of the global transcriptional landscape, which allows an affordable and accurate approach for gene expression quantification and differential gene expression analysis between multiple groups of samples. RNA-Seq can identify novel and previously-unexpected transcripts without the need for a reference genome, allowing de novo assembly of new transcriptome that is not previously studied before. It also enables the discovery of novel gene structures, alternatively spliced isoforms, gene fusions, SNPs/InDel, and allele-specific expression (ASE).
Advantages of RNA-Seq
- Quantitative and precise measurements of RNA molecules at a single base-pair resolution
- Discovery of novel transcripts, splice variants, and gene fusions
- Application to any species, no matter whether reference genome is available
- A comparable or lower price than many other methods
RNA-Seq Workflow
SeqInsight combines both Illumina HiSeq and PacBio systems to provide a fast and accurate RNA-Seq and bioinformatics analysis for any species. The general workflow for RNA-Seq is outlined below.
Service Specification
Sample requirements and preparation
• RNA amount ≥ 2 μg, RNA concentration ≥ 50 ng/μl, OD260/280=1.8~2.0 • All RNA samples are validated for purity and quantity
Sequencing
• Regular: Illumina HiSeq PE150 • Differential gene expression study: Illumina HiSeq 50 • Full-length transcript: PacBio SMRT • More than 80% of bases with a ≥Q30 quality score
Bioinformatics Analysis
We provide customized bioinformatics analysis including: • Statistics of sequencing depth and coverage • De novo assembly, and reference genome mapping • Gene annotations and gene expression levels • Prediction of novel RNA and identification of variants • Test for differential gene expression • Assessment of allele-specific expression • Identification of expression quantitative trait loci (eQTLs)
Analysis pipeline
SeqInsight provides full RNA sequencing service package including sample standardization, library construction, deep sequencing, raw data quality control, genome assembly, and customized bioinformatics analysis. We can tailor this pipeline to your research interest. If you have additional requirements or questions, please feel free to contact us.