Total RNA-Seq (Whole Transcriptome) sequencing captures a broader range of gene expression changes and enables the detection of novel transcripts in both coding and non-coding RNA species. Ribo-Zero ribosomal RNA reduction chemistry is used in place a poly-A tail selection which efficiently removes ribosomal RNA (rRNA) This process minimizes ribosomal contamination and optimizes the percentage of reads covering RNA species of interest.
LC Sciences provides a one-stop solution (i.e. from sample to data) for Total RNA-Seq using the latest in RNA-Seq technology. The transcriptome profiling and analysis services by LC Sciences are comprehensive and provide the most complete picture of RNA content in your samples. We can help you set up and conduct a high-quality, well-controlled Total RNA-Seq experiment based on the latest deep-sequencing technologies.
- Library preparation – ribosomal RNA depletion method
- Illumina Hi-Seq 2000 – 2 x 100bp paired-end sequencing run
- Advanced bioinformatics package
Our Comprehensive Service
LC Sciences provides a transcriptome profiling service using the Illumina high-throughput sequencing technology which enables comprehensive coverage, highly sensitive and specific discovery and profiling of all forms of RNAs in your sample.
This service is comprehensive – from sample to data, providing advanced technology and years of experience in efficient discovery and profiling of RNAs.
Our comprehensive profiling service includes:
- Sample QC
- Sample preparation (ribosomal RNA depletion, adapter attachment, reverse transcription)
- Library preparation (single molecule clonal amplification)
- High-throughput sequencing (RNA-Seq method, Illumina Next-Gen sequencing technology)
- Bioinformatics analysis
- Customer data report
Data Analysis Includes:
- Illumina base-calling and analysis
- Raw sequencing data – 100bp paired-end reads – either FASTA-A or FASTA-Q files with base-calling quality scores
- LC Sciences quality filtered data reduced to mappable reads, a list of unique sequences and their copy numbers
- De novo assembly (if applicable)
- Alignment of RNA-Seq reads to customer specified reference genome
- Identification and construction of splice-junctions
- Report of known transcripts with annotation and abundance
- Report of novel transcript with abundance
- Identification of alternate splicing and report of isoform abundance
- Test for differential expression at gene level and transcript level. (biological replicates required – see Technical Note – Biological Replicate Considerations – Differential Expression Analysis)
- GO and KEGG annotation and enrichment analysis (optional)
- Non-coding RNA information – map to genome, alignment, limited annotation and expression info
- Optional circular RNA (circRNA) analysis – although the biological function of most circular RNAs is still unclear, some have shown potential as gene regulators.
- Customer data report – includes a summary of methods and all statistical analysis