Carefull processing of your sample material performed by experienced hands in optimal facilities
To prepare your material for deep-sequencing, three essential steps are required in the laboratory. At omiics, we have the facilities and experience to conduct all three for you.
RNA EXTRACTION
Isolation of RNA from cells, tissues or biofluids
LIBRARY PREPARATION
Prepare RNA for sequencing choosing the optimal kit
To sequence your sample, we start with RNA isolation.
The optimal procedure depends on whether you are interested in sequencing total/long RNA (mRNA, lncRNA, circRNA) or small RNAs (miRNA, tRNA), and the sample material.
Sample material
omiics has experience with a vast array of biological sample material. Examples include:
Sample quality
We specialize in low-input samples (e.g. cell-free material) and have successfully sequenced samples using input as low as 250 pg RNA. For standard setups, we recommend input of 500 ng (total RNA) to 1 μg (small RNA) for sequencing.
Sample quantity
RNA is a highly unstable molecule. It is crucial to handle samples in an RNAse-free environment and keep samples below 4 °C at all times.
Unique material comes in limited amounts and may for different reasons be of low quality.
Occasionally, samples isolated using methods essential for addressing novel questions may damage the RNA. This may hinder some analyses but not all.
In omiics, we have extensive experience with getting information from even low quality samples.
Lets talk about what we can extract from your precious samples
Today, a wide selection of library preparation kits available and the best choice depends on the input quality and quantity, as well as the type of end-analysis needed.
Make it easy for yourself - Let omiics choose the optimal reagents for your setup.
Deplete the many or enrich the few?
Ribosomal RNA (rRNA) comprises a significant proportion (~90% or more) of all the RNA in total RNA samples. While the rRNA will not directly cause harm to the result, removing rRNA or enrichment of mRNA prior to generating libraries provides benefits by lowering sequencing costs and improving mapping statistics.
Unique molecular identifyer (UMI)
A speciality of omiics is sequencing ultra-low input samples down to as low as 250pg RNA.
At ultra-low RNA concentration, the sample becomes more sensitive to PCR-associated bias. By including an additisonal molecular tag during library preparation, the unique molecular identifier, UMI, enables you to get high-resolution reads, to facilitate accurate detection of true variants.
For each step of the process in the laboratory we have strict quality control. If for some reason, a sample fail to pass our standards, we will contact you and discuss the options.
RNA quality
The quality and concentration of extracted RNA is analyzed by Qubit and Bioanalyser 2100.
Based on concentration and the RIN value it is decided if the material is appropriate for the contracted sequencing design.
Library quality
Library quality is determined using the Bioanalyzer 2100, and accurate concentration of sequencable library is estimated by qPCR.
Once passed, the next step is sequencing
What to expect in your report from omiics
Customized bioinformatics to get the most from your data
NGS service provider with a focus on customer needs. We deliver flexible solutions to your specific demands.
Contact us for a non-binding discussion about your next project.
