Do not neglect the small ones
Discover new small RNAs and analyse the expression of known small RNAs.
Small RNAs constitute a family of RNA species that plays a central role in regulating gene expression in many organisms. With the enormous depth reached by next generation sequencing technologies, you can get a comprehensive view of the small RNAs in your samples.
Sequencing small RNA is done by preparing a short insert cDNA library for each sample and subsequently sequencing these libraries using Illumina NextSeq 500 or MiSeq technology. To sequence several small RNA samples in one channel, we offer library preparation with individual indexes.
Typical projects are:
- Differential expression analysis of small RNAs
- Identification of novel small RNAs
- Identification of target molecules of novel miRNAs
We accept isolated RNA, microbial cultures (non‐pathogenic), animal/plant tissues, etc
- RNA: Submit DNase treated RNA of approximately 3-5 µg of high quality with Nanodrop A260/280 ratio >1.9; A260/230=2.0-2.2; and at a concentration of ~100 ng/µl (dissolve in nuclease free water). Supplied RNA should be shipped in dry ice to the Eurofins facility.
- Plant/Human/Animal/ Insect Tissue: Tissues of 2-4 gm should be provided in RNA later or snap frozen tissue without any storage solution shipped in dry ice.
- Microbial non-pathogenic culture: Snap frozen pure isolated culture (Bacteria / Yeast / Fungi) should be shipped in dry ice.
Sample Quality Control:
- NGS grade High quality RNA will be isolated from the supplied sample using commercially available kits.
- The qualification and quantification of RNA will be carried out by denatured agarose gel electrophoresis and NanoDrop/Qubit Fluorometer analysis.
- RIN value estimation will be carried out using Agilent 4200 Tape Station.
- Small RNA Seq library will be prepared by using TruSeq small RNA library preparation Kit.
- Library validation will be carried out using High Sensitivity D1000 Screen tape using Agilent Tape Station.
- Sequencing will be carried out on Illumina NextSeq 500 platforms with 1 x 75 single end chemistry.
- Generation of required amount of data.
- Sequence data (base call files or bcl files) generated from the sequencer are de-multiplexed if required and converted to FASTQ files.
- Raw data will be available for download as a compressed archive of FASTQ files for each sample.
- Raw data can be post processed upon request.
- Comprehensive compiled report and data set in Pen Drive or Hard Disk.
- 5-7 weeks after arrival of your samples and all necessary information. It depends upon data size, scope, technology selected, number of samples and complexity of the project. The TAT is offered expecting no biological or technical difficulties for processing of all project samples.