The incredible power of RNA-Seq was initially applied primarily for complex whole transcriptome applications such as discovery of rare genes, splice junctions and gene fusions or for study of novel and less well-characterized organisms. However, as the technology is improving and costs are decreasing, RNA sequencing is now suitable to be used for simple RNA profiling (sample comparison) as well.
RNA-Seq in fact holds several advantages over microarrays for gene expression analysis applications:
- Expression profiling is truly genome-wide – there is no need for probes based on prior sequence information.
- Powerful RNA-Seq technology reads every base of every transcript – this is not TAG profiling.
- Achieve digital transcript expression analysis – accurate quantitation of expression levels over 5 orders of magnitude of transcript abundance.
Our Comprehensive Service
LC Sciences provides a a one-stop solution (i.e. from sample to data) gene expression profiling service using the Illumina next-generation sequencing technology which enables comprehensive coverage, highly sensitive and specific profiling of mRNAs 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 (poly(A) selection, 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 – 50bp single-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
- Genomic mapping of quality sequencing reads
- Test for differential expression at gene level. (biological replicates required – see Technical Note – Biological Replicate Considerations – Differential Expression Analysis)
- GO and KEGG annotation and enrichment analysis.
- Customer data report – includes a summary of methods and all statistic analysis