Test the library quality before sequencing
Prepare RNA for sequencing
Isolation of RNA from cells, tissues or biofluids
LABORATORY PREPARATION OF MATERIAL FOR SEQUENCING
To prepare you 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.
OTHER LABORATORY SERVICES
At the omiics laboratory unit, we perform additional services as well, and the selection keeps growing.
SAMPLE MATERIAL SPECIFICATION
omiics has versatile experience with sample material from tissue, cells, biofluids and powders. Examples include muscle and brain tissue, mammalian cells, exosomes, blood, CSF, bile, saliva and food powders.
We specialize in low-input samples and have successfully sequenced samples using input as low as 250pg RNA. For standard setups, we recommend input of 500ng (total RNA) to 1ug (small RNA) for sequencing.
RNA is a highly unstable molecule. It is crucial to handle samples in an RNase-free environment and keep samples below 4C at all times. As a standard, RNA should have a RIN value above 7 for optimal sequencing. Partially degraded RNA samples can also be sequenced.
TOTAL LONG RNA
Ribosomal RNA (rRNA) comprises a significant proportion (~90% or more) of all the RNA in total RNA samples. Depleting rRNA from total RNA prior to generating libraries provides benefits by lowering sequencing costs and improving mapping statistics.
To perform sequencing of your sample, we initially isolate RNA. The optimal procedure is depends on whether you are interested in:
TOTAL LONG RNA & SMALL RNA
Today there is a wide selection of library preparation kits available and which one is the best choice depends on both the input quality and quantity, as well as, the type of end-analysis needed. Let omiics choose the optimal reagents for your setup.
Before we make the final decision to sequence, we check the quality of the finished libraries using the Bioanalyzer technology.
The concentration is determined with high accuracy using a specialized qPCR protocol
The Nanostring technology constitutes a valuable approach to screening the expression level of selected candidates. It can serve as a simplified alternative to NGS, or it can be used to validate the expression of interesting candidates identified by NGS to improve the impact of the findings. Nanostring is often viewed as a time-saving alternative to qPCR validation.
ADDITIONAL METHODS AND EQUIPMENT
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