small RNA sequencing

small RNA sequencing services

For untargeted analysis of microRNAs and other small RNAs, next-generation sequencing (NGS) is the platform of choice, as it enables unbiased genome-wide characterization of small RNAs in biological matrices. At TAmiRNA we are fully dedicated to microRNA and small RNA NGS analysis. Our customers benefit from the many years of experience our team has in microRNA and small RNA biomarker research and development. As we are working in biomarker research, in addition to working with cells and tissues we have experience with low-input RNA samples such as microdissected tissue and biofluids such as serum, plasma, urine, saliva, CSF, and synovial fluid as well as cell culture supernatant. Recently, we have expanded our service to extracellular vesicles and protein complexes. Our NGS services go hand-in-hand with our RT-qPCR services to further verify and validate NGS findings.

small RNA sequencing
small RNA sequencing

benefits

Our service includes total RNA extraction, small RNA sequencing library preparation, next-generation sequencing, data analysis, and reporting. Depending on customer experience we can focus on individual tasks or provide a full service.

Our small RNA sequencing workflow has undergone analytical validation. With best-in-class precision and accuracy, we can ensure the highest data quality. By using a novel and proprietary spike-in calibrator we can transform relative counts into absolute concentrations.

We provide ready-to-use data that can directly be integrated into reports, publications, and presentations.

We have conducted small RNA sequencing analysis with samples from the following organisms:

  • Human
  • Non-human primates
  • Mouse
  • Rat
  • Pig
  • Cow
  • Horse
  • Chicken
  • Dog
  • C. elegans
  • D. melanogaster
  • Chinese hamster ovary cells

For other organisms data analysis can be implemented upon request.

We offer MiSeq, iSeq, HiSeq2500, NextSeq550 and the NovaSeq. Thus, we manage to find a cost-effective solution for small and large projects.

sample requirements

Cells: > 10 cells

Tissues: we can process everything from laser microdissected compartments (> 10,000 µm2) to frozen or fixated tissue blocks.

Biofluids: serum, plasma, urine, saliva, CSF, synovial fluid. Preferred input is 100-200 µl but can be optimized to meet bottlenecks.

Extracellular vesicles: depends on the purification method. We prefer to discuss individually.

small RNA sequencing

service description

Small RNA sequencing includes several steps, which can all be performed at TAmiRNA:

  • Total RNA extraction: we have protocols for cells, tissues, biofluids and EVs. Leftover total RNA can be returned to customers for follow-up experiments.
  • Total RNA quality control: concentration and integrity using highly sensitive and specific fluorescence based methods
  • small RNA library preparation: we prepare high quality libraries using a circularization protocol. This reduce adapter ligation bias compared to other methods. In addition, we have developed a proprietary spike-in calibrator that is added to the library preparation and enables absolute quantification.
  • NGS analysis: using a wide-range of Illumina instruments (HiSeq, NextSeq, NovaSeq) we can pick the most efficient solution for each project.
  • Data analysis: we provide everything from raw data to fully analyzed data reports depending on customer needs.

reference projects

Biotransformation of the Mycotoxin Zearalenone to its Metabolites Hydrolyzed Zearalenone (HZEN) and Decarboxylated Hydrolyzed Zearalenone (DHZEN) Diminishes its Estrogenicity In Vitro and In Vivo.

Fruhauf S, Novak B, Nagl V et al. Toxins (Basel). 2019 Aug 20;11(8). pii: E481. doi: 10.3390/toxins11080481

MicroRNA Expression Profile Changes after Cardiopulmonary Bypass and Ischemia/Reperfusion-Injury in a Porcine Model of Cardioplegic Arrest.

Kiss A., Heber S., Kramer AM. et al. Diagnostics (Basel). 2020 Apr 21;10(4):240. doi: 10.3390/diagnostics10040240.

Anti-CD3 Antibody Treatment Reduces Scar Formation in a Rat Model of Myocardial Infarction.

Wernly B., Paar V., Aigner A. et al Cells. 2020 Jan 25;9(2):295. doi: 10.3390/cells9020295.

Combined proteomics/miRNomics of dendritic cell immunotherapy-treated glioblastoma patients as a screening for survival-associated factors.

Erhart F., Hackl M., Hahne H. et al. NPJ Vaccines. 2020 Jan 16;5:5. doi: 10.1038/s41541-019-0149-x. eCollection 2020.

MicroRNA levels in bone and blood change during bisphosphonate and teriparatide therapy in an animal model of postmenopausal osteoporosis.

Kocijan R., Weigl M., Skalicky S. et al. Bone. 2020 Feb;131:115104. doi: 10.1016/j.bone.2019.115104. Epub 2019 Nov 1.

MicroRNAs in porcine uterus and serum are affected by zearalenone and represent a new target for mycotoxin biomarker discovery.

Grenier B., Hackl M., Skalicky S. et al. Sci Rep. 2019 Jun 28;9(1):9408. doi: 10.1038/s41598-019-45784-x.

Predicting Postoperative Liver Dysfunction Based on Blood-Derived MicroRNA Signatures.

Starlinger P., Hackl H., Pereyra D. et al. Hepatology. 2019 Jun;69(6):2636-2651. doi: 10.1002/hep.30572. Epub 2019 Apr 10.

Small extracellular vesicles and their miRNA cargo are anti-apoptotic members of the senescence-associated secretory phenotype.

Terlecki-Zaniewicz L., Lämmermann I., Latreille J. et al. Aging (Albany NY). 2018 May 19;10(5):1103-1132. doi: 10.18632/aging.101452.

A signature of 12 microRNAs is robustly associated with growth rate in a variety of CHO cell lines.

Klanert G., Jadhav V., Shanmukam V. et al. J Biotechnol. 2016 Oct 10;235:150-61. doi: 10.1016/j.jbiotec.2016.03.022. Epub 2016 Mar 16.

Next-generation sequencing analysis of circulating micro-RNA expression in response to parabolic flight as a spaceflight analogue

Jirak P., npj Microgravity volume 6, Article number: 31 (2020)

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small RNA sequencing services