1. Library preparation – ribosomal RNA depletion method
  2. Illumina Hi-Seq 2000 – 2 x 100bp paired-end sequencing run
  3. Advanced bioinformatics package

Next-Gen DNA Sequencer

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

  • 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