How Our Services Work
Find a workflow that best meets your needs from sample submission to data analysis
Each project begins with a free consultation with an expert to design an experimental setup that best meets your needs and budget. Following this, we will complete a detailed sample-submission form, making sure that all experimental details and subsequent analyses are clearly defined.
Please contact us for further product information, or technical questions.
The How Our Services Work is intended for molecular biology applications. This product is neither intended for the diagnosis, prevention, or treatment of a disease, nor has it been validated for such use either alone or in combination with other products.
RNA sample submission
High-quality samples are important for accurate RNA sequencing. At the initial consultation, we offer recommendations on suitable extraction and clean-up methods. Alternatively, you can take advantage of our expertise and submit your samples to our RNA isolation service.
mRNA or whole transcriptome
We recommend that you send us 260 ng total RNA, but we get high-quality results with as little as 100 ng total RNA + 60 ng for QC.
For ultra-low mRNA NGS, RNA isolation is included in the project and must be performed by us as we have developed an optimized NGS workflow with proprietary steps including unique RNA isolation and QC protocols specifically optimized for samples with low RNA content. Ultra-low mRNA sequencing is ideal for small amounts of cells or tissue including fine needle biopsies, LCM samples and sorted cells. We recommend that you send us a minimum of 100 cells to get high-quality results.
For miRNA-seq in serum and plasma, RNA isolation is included in the project and has to be performed by us. Our optimized NGS workflow with proprietary steps, including unique RNA isolation and QC protocols, is specifically optimized for miRNA sequencing from serum/plasma. Although we get high-quality results from as little as 250 µl, we recommend sending 500 µl of serum/plasma. We accept human serum/plasma samples, and samples from other species or other biofluids (urine, CSF, saliva, etc.) upon request.
We offer to isolate RNA from your samples for sequencing. We perform high-quality RNA isolation from a range of sample types including cells, tissues, FFPE sections and blood. We also perform RNA isolation from minute amounts of sample as part of our Ultra-low mRNA Sequencing Service.
Isolation of RNA from biofluids is performed using QIAGEN kits and protocols developed specifically for this sample type.
RNA sample quality control (QC)
After receiving your RNA samples, we will determine the integrity and quantity of each sample using a Bioanalyzer/TapeStation and Nanodrop instruments and possible contaminations are assessed for each sample. You will receive a report with the results of these analyses prior to the library preparation and sequencing. We also perform the QC prior to any RT-reaction for downstream qPCR profiling.
Following RNA isolation, we will perform RNA QC using qPCR-based methods optimized specifically for low RNA content samples. The RNA input amount is normalized by quantification of GAPDH and ß-actin using qPCR. You will receive a report with the results of these analyses prior to the library preparation and sequencing.
For biofluid samples, QC of the RNA is performed by qPCR, using a unique combination of RNA spike-ins and endogenous miRNAs to detect outlier samples and monitor RNases, RNA isolation efficiency, hemolysis and presence of inhibitors that may affect library preparation. You will receive a report with the RNA-QC results prior to the library preparation and sequencing.
After QC, we design libraries according to the type of sequencing to be performed. For mRNA sequencing, we enrich poly-A mRNA molecules from total RNA using an Oligo-dT magnetic bead-based system. For whole transcriptome sequencing, we perform ribosomal RNA depletion (through the use of a biotin-streptavidin bead-based system). The resulting RNA is fragmented and converted into mRNA or whole transcriptome cDNA NGS libraries. A Bioanalyzer/TapeStation is used to perform QC of the libraries.
We generate libraries and select size using a bead-based size selection of 15-40 nt. Library QC is performed using a Bioanalyzer/TapeStation.
We offer mRNA and whole transcriptome sequencing using the Illumina platform. We use both NextSeq 500 and HiSeq 2500 instruments, enabling us to cater for a range of project sizes. For standard projects, the sequencing parameters are 1 x 75 bp, 30 M reads per sample or 2 x 75 bp, 60 M paired-end reads per sample. However, the sequencing parameters can be tailored to your specific requirements. Please contact us for further information.
We offer 2 types of miRNA sequencing using the Illumina platform:
Biofluid miRNA sequencing is performed using Illumina instruments, 1x 75bp reads, 10M reads per sample.
Analysis and interpretation for mRNA and whole transcriptome NGS projects
Comprehensive data analysis, including statistical analysis is provided using our unique NGS data analysis pipeline:
Report and final consultation
The final report is delivered by our cloud-solution called XploreRNA and contains:
mRNA or whole transcriptome and miRNA: 8–10 weeks
Ultra-low mRNA amounts or biofluids: 8–10 weeks
After you have received your report, we arrange a follow-up discussion to answer any questions you may have about your report and discuss the significance of your results and next steps. We can also help you design appropriate miRCURY LNA or RT2 qPCR assays to validate your NGS data.
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