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.
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.
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.
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.
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.