omiics | Small RNA

SMALL RNA SEQUENCING

Small RNAs

miRNA, tRNA fragments (tiRNA), siRNA, piRNA

and more

Precise quantification of small RNA

MicroRNA (miRNA) is known to as a vital control mechanism. Each miRNA can control hundreds of messenger RNAs (mRNA), and a mRNA is most often under the control of several miRNAs, resulting in a finely balanced, intricate network of control.

Gaining insight into small RNA expression signature in cells, tissue and biofluid of interest can have great impact on biomarker discovery, detection of treatment targets and insight into mechanisms involved.

While each project is unique and consequently the optimal solution varies according to the settings, some elements will be fairly consistent across most experiements.

A complete omiics standard project for small RNA sequencing include:

The project starts with a meeting with a NGS-scientist, where your research

and material is discussed and the optimal startegy is designed.

RNA extraction Tissue, cells, exosomes, plasma, bioliquids, FFPE
Library preparation

- 18-40bp insert cDNA library

- Multiple kit options available

Stranded sequencing*

LABORATORY

* Non-stranded sequencing is available, however, gives you less valuable data and while historical cheaper, the current prices are identical.

Platforms: NextSeq500 as standard
≥10 Million reads/sample as standard
75bp SE

SEQUENCING

SE: Single-end

Quality control - scoring of reads and adaptor removal, incl. assessment of sequencing statistics
Mapping to known genome/transcriptome (alignment) including sequencing assessment/alignment statistics
Distribution of small RNA reads across RNA species incl. microRNA, rRNAs, tRNAs, snoRNAs and more
Differential expression analysis and cluster analysis (Gene expression annotation, expression pattern analysis, Hierarchical clustering analysis, PCA analysis, quantitative expression profiles)

BIOINFORMATICS

Tables (.excl*)
Publication-grade figures (.TIFF*)
Written report summarizing the procedure and results done by omiics
Follow-up meeting with PhD-level NGS-scientist from omiics discussing results, suggestions for interpretation and input on possible next step(s)

REPORT AND DISCUSSION

* Other formats are also available if requested

Additional

options

below

More info on input material

or Species?

Additional Bioinformatic Analysis for small RNA sequencing

Target mRNA prediction
Pathway analysis (Gene ontology (GO) analysis, Pathway enrichment analysis KEGG
Interaction analysis between differentially expressed mRNAs and differentially expressed miRNAs.
Length distribution of small RNAs
Prediction of novel miRNAs and their secondary structures by mirDentify and miRDeep-2 from
unannotated small RNAs
Family analysis of known miRNAs
Predictive biomarker modelling by lasso-regression

Customized analysis - contact us to discuss your needs