Experts in RNA-seq
Differential expression analysis by RNA-seq
RNA-seq is a powerful, cost-effective tool of great interest in several life science areas. RNA-seq allows us to obtain a large number of reads from the transcriptome. These reads are further analysed using high-throughput bioinformatic methods. The most common application of RNA-seq is to detect differentially expressed genes under given environmental stressors.
What you receive
With our standard service you receive a summary of the methods followed, the raw high-throughput sequencing files, and the results of the bioinformatic analyses including demultiplexing, pre-processing and quality control of the raw data, read alignment against the reference genome, gene expression quantification, annotation overlap statistics, and detection of differential gene expression among conditions. If you require additional analyses, please let us know and we will do our best to meet your needs.
What we need
We can start working by using either your tissue samples, or RNA which has already been extracted.
If you are sending tissue samples, please make sure that they have been properly preserved for RNA isolation (liquid nitrogen, RNAlater® or other suitable RNA preservation agents).
If you are sending RNAs which have already been extracted, please note that we ideally require a minimum amount of 2 μg of high-quality total RNA in nuclease-free water (concentration >40 ng/μL) per sample, as measured by the Agilent 2100 Bioanalyzer or equivalent. RNA should be shipped in dry ice to prevent it from degrading.
Please note that, depending on the specifics of your project, these requirements may vary. Please contact us for the specific requirements of your project.
How we work
Step 1: we isolate RNA from your samples. We have adapted different RNA isolation protocols depending on the starting biological material.
Step 2: we purify the RNA fraction you are interested in (total RNA, mRNA, small RNAs, noncoding RNAs, circular RNAs) and prepare cDNA libraries using the kits recommended by Illumina.
Step 3: we sequence the libraries using the Illumina HiSeq technology. Millions of reads per library are obtained in this step.
Step 4: we analyse the high-throughput sequencing data in collaboration with our partner ecSeq.
For your convenience, we can carry out the entire project or only those steps you require. As a norm, steps 2 and 3 should be ordered jointly. Please contact us for further information and pricing.