RNA-Seq to reveal a snapshot of RNA presence and quantity from a genome at a given moment in time

RNA-seq can provide a profile of RNA presence, quantity, and sequence at a given moment in time or under specific conditions. Applications include:

  • Gene Expression, identification of alternative gene spliced transcripts
  • Differential Gene Expression comparing control and test conditions for example
  • Pathway analysis to identify metabolic pathways affected under test conditions
  • Single Nucleotide Variation Discovery
  • Post Transcriptional Variations
  • Fusion Gene Detection
  • Transcriptome assembly of novel genomes

PolyA RNA-seq

Selective enrichment of poly-adenylated transcripts for cDNA preparation. Strand bias is not conserved.

Human genetic disorders

Whole Transcriptome

Pre-enriched RNA submission OR selective rRNA depletion of total RNA is followed by cDNA preparation allowing detection of lncRNA as well as mRNA (≥100nt). Strand bias is not conserved.

Human genetic disorders

Strand-specific library preparation

Ligation of unique adapters to the 5′- and 3′-ends of RNA fragments allows discrimination of the template DNA strand. This can be applied to either polyA or whole transcriptome projects.

Human genetic disorders

smRNA-seq/miRNA-Seq

Similar to strand-specific library preparation, with size selection to limit insert size of 19-21nt.

Normalized libraries

Treated with duplex-specific nuclease (DSN), resulting in under-representation of highly expressed transcripts, allowing detection of less abundant transcripts. Not suitable for experiments that need to retain expression profiling information.

Custom Target RNA-Seq

Please inquire with a list of target of interest.

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Sample requirements

Total RNA (1-5 micrograms) or any source of biological samples (cells, tissues, FFPE, blood). Experienced staff at myGenomics can extract RNA from variety of sample types. Min. concentration: 25 ng/µl; OD 260/280 ≥ 1.8, RIN ≥ 8

Turnaround time:

Sample-Report: 20-30 days

Results

  • Raw data (FASTq files) and if analysis requested:
  • Gene expression report, FPKM values,
  • Differential gene expression report (CuffDiff, TopHat),
  • Pathway identification
  • Single Nucleotide Variation discovery
  • Fusion Gene Detection

Additional deliverables

Initial sample QC, cDNA Library prep QC, Fragmentation QC, Sequencing reads QC, Coverage and FastQC plots.

Introduction to RNA-Seq analysis