Technology & Service
For high throughput sequencing the unit employs the Illumina technology that uses massively parallel sequencing to generate several billion bases of high quality nucleotide sequences per run. To this end DNA molecules are attached to a surface, amplified and the complementary DNA strand re-synthesized by consecutive incorporation of fluorescent reversible terminator deoxyribonucleotides (a technology referred to as sequencing-by-synthesis). Surface-images are analyzed to generate high-quality sequences. Using the Illumina sequencing technology maximum read length of 100 bp (HiSeq2500, high-output mode), 150 bp (HiSeq2500, rapid-run mode) as well as 300 bp (MiSeq) is feasible. Furthermore paired-end sequencing can be applied as well as barcoding (indexing, multiplexing) to increase the number of parallel analyzed samples.
For high output as well as ultra-fast sequencing the facility operates the most recent Illumina sequencers, a HiSeq2500® and a MiSeq®. Furthermore a robust instrumentation infrastructure is in place for standardized and quality controlled sample preparation. This includes a Covaris S220, an Agilent Bioanalyzer, a Qubit Fluorometer as well as liquid handling stations for automated chromatin immunoprecipitation and library preparation.
To manage the huge data flow generated by the facility, we utilize a large data center and computer cluster for storage, analysis and visualization.
- Preparation of multiplexed libraries for various applications:
- RNA-Seq (Poly-A, Ribo-, stranded)
- gDNA-Seq and Amplicon-Seq
- Cluster generation and sequencing using the HiSeq2500 and MiSeq instrument