Profiling of community-wide gene expression (RNA-seq)
Analyze the function and activity of the complete set of transcripts (RNA-seq) from environmental samples. While metagenomics tells us which microbes are present and what genomic potential they have, on the other hand metatranscriptomics tells us about their activity: the genes that are highest expressed in a specific microbial environments.
- Submit 10-20 µg of DNA free RNA, Nanodrop A260/280 ratio >1.9; A260/230=2.0-2.2; and at a concentration of ~100 ng/µl
- Or soil, plant/ human microbiome, water or sludge samples, shipped in cool packs to eurofins facility.
- NGS grade RNA isolation will be carried out using commercially available RNA isolation kit.
- Further quality checking will be carried out using denatured agarose electrophoresis and NanoDrop. Quantification will be carried out using Qubit 3.0 fluorometer.
- RIN value estimation will be carried out using Agilent tape station.
- Ribosomal RNA depleted / mRNA enriched RNA Seq library will be prepared using illumina TruSeq RNA v2 LT kit.
- Library validation will be carried out using High Sensitivity D1000 Screen tape using Agilent Tape station kit.
- Sequencing will be carried out on illumina NextSeq 500 using 2 x 75 or 2 x 150 bp paired end sequencing chemistry to generate 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.
- 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.