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RNA Sequencing

RNA Sequencing (RNA-Seq) is an emerging technology that is transforming the landscape of current and future gene-expression studies. While traditional methods such as microarray and comparative genomic hybridization have been dominant over the last decade, RNA-Seq is expeditiously replacing these technologies through delivery of data that is proving to have higher sensitivity and accuracy. The RNA-Seq service offered at Microgem produces high quality sequencing data and accurate gene expression level estimation to facilitate your next big discovery.

Key features for RNA-Seq Service:

  • Sequencing Platform
    •  Illumina
  • Sequencing Scale
    • 8 million reads for rapid expression analysis
    • 40 million reads for detecting alternative splice forms
    • 80 million reads for identifying low-abundance coding and non-coding transcripts
    • Higher coverage available up to 400 million reads per sample
  • Starting Material
    • 0.1µg – 4µg of Total RNA
    • 10-100ng of Poly-A enriched mRNA
    • 10-100ng of rRNA depleted RNA
  • Sequencing Type
    • 75 bp single end or paired end sequencing
    • Longer read lengths available upon custom request
  • Library Type
    • Strand-specific
  • Bioinformatics Analyses
    • FastQC on raw sequencing data (included)
    • Read alignment and estimation of gene expression (included) Differential gene expression analysis
    • Functional annotation
  • Turn-around Time
    • 2-4 weeks from passing sample QC plus 1-2 weeks for analysis
  • Data Storage
    • 3 months

Standard bioinformatics analyses

Microgem is quick to adopt the state of the art RNA-seq analysis methods that is orders of magnitude faster with higher sensitivity and specificity than current popular choices of TopHat2, Cufflinks, and Cuffdiff.

Exploratory Bioinformatics Analyses

At Microgem, we help our collaborators to jump start their data exploration by performing various statistical analyse including sample correlation, PCA, and differential gene expression analysis. The figures and tables are provided to quickly identify key genes or samples that will be of interest to you. Using published sample data, we show a few representative figures below.

Deliverables

  1. Sequencing results in FASTQ format
  2. Expression level estimation table (FPKM and TPM)
  3. Differentially expressed gene table (If applicable)
  4. Functional annotation table (If applicable)
  5. QC and bioinformatics analysis report