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Next-Generation-Sequencing (NGS) is a high-throughput method for massive, parallel sequencing of millions of nucleotide fragments.
A NGS experiments needs a carefully evaluated study design, good quality source material (DNA or RNA), a so-called library preparation,
the sequencing run to generate big datasets and bioinformatic / statistician expertise to extract meaningful biological results from these data.
In our institute, we offer to do the library preparation process as well as the NGS sequencing on the basis of a scientific collaboration.
We can also set up contact with bioinformaticians helping
with experimental planning as well as analysis.
Currently, the fleet of NGS sequencing devices in our institute comprises the Illumina
NovaSeq X-Plus,
NovaSeq 6000,
NextSeq 1000,
MiSeq i100 and the old
MiSeq systems
and a PacBio Revio and Sequel IIe systems.
The Illumina MiSeq and its successor MiSeq i100 is a small benchtop sequencer with a maximum output of 25 M paired-end reads and 25 Gigabases.
Due to its low throughput and rapid turnaround time, the MiSeq is recommended for targeted re-sequencing (small gene panels, e.g. BRCA),
amplicon sequencing (e.g. 16S) and sequencing of small genomes (e.g. bacteria) as well as for QC of larger, expensive runs to determine
loading concentrations, pooling ratios and/or success of library preparation.
See output specifications of the MiSeq
and MiSeq i100.
The Illumina NextSeq is a small high-throughput benchtop sequencer with a maximum output of 400 M paired-end reads and 240 Gigabases of data.
With the choice of a variety of different Flowcells, the NextSeq is a highly flexible device and can be used for Exome sequencing, transcriptome
sequencing and Panel-Sequencing. Because of its rapid turnaround time, we recommend this device for urgent samples and projects with low sample numbers.
See output specifications of the NextSeq 1000 here.
The NovaSeq 6000 is Illuminas previous flagship model. With a large variety of different Flowcells (SP, S1, S2, S4) offering different read-length
and data outputs, the NovaSeq is highly flexible device. With up to 10 billion reads and 3000 gigabases output the NovaSeq is a high-throughput sequencer,
suitable, e.g. for exomes, genomes and transcriptomes. 4 lanes can be loaded individually. We still sequence ongoing experiments on the NovaSeq 6000 for
scientific consistency, but would recommend to move new experiments to the NovaSeq X.
See output specifications of the NovaSeq 6000 here.
The NovaSeq X-Plus is our latest addition to the short read sequencer lineup. The availability of different flowcells (1.5B, 5B, 10B, 25B) makes
it a highly flexible device. A Flowcell generates up to 25 billion reads and 10 terabases of output, making it the number one choice for all big data projects.
8 Lanes can be loaded individually (except on the 1.5B with 2 lanes). Maximum read length is 2x300 bp paired-end.
See output specifications of the NovaSeq X-Plus here.
The PacBio Revio is a long-read sequencer of the third generation, producing reads with read-lengths up to 20 kilobases and an output up
to 120 Gb per Flowcell. With the circular-consensus-sequencing technology, read accuracies of 99.99 % can be achieved. The Revio is
recommended for de novo genome projects and structural variant detection as well as full length transcript RNAseq (IsoSeq).
Because the Revio can sequence native DNA, 5mC and 6mA Methylation-Sequencing is also included. In addition, multiplexed long
amplicon sequencing (e.g. full lengths 16S) is possible.
See specifications of the Revio here
and information on the special requirements of third generation sequencers here.