In case of specific requirements, we can provide sequencing using also other sequencing platforms
Last update: November 2 2021. For those interested in NGS technology we recommend our regular two-day course! More information here.
Currently we offer processing of your samples using following technologies and instruments. Given the rapid technological developments, it is possible that despite all efforts, the information provided below is not 100% up to date.
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NovaSeq6000 (Illumina)Instruments by Illumina have at the moment the highest accuracy of reading with error rate <0,001. We can offer instruments and chips having various outputs of sequencing data in terms of read length and amount (sequencing capacity). Using the proper settings and data allocation (see ShareSeq) it is possible to ensure economic use of the device meaning with one type of technology, experiments with different sequencing capacity requirements can be realized. It is possible to carry out a pilot experiment with a lower capacity and then design a larger scale experiment according to the results. All Illumina instruments use the sequencing chip called flow-cell, in higher-capacity models with several separate lanes. NovaSeq6000 flowcell S4 (total capacity 3 Tb) These lanes can be considered as separate units for sequencing. Thus, multiple samples can be sequenced in one sequencing run without indexing. Using indexing, it is also possible to analyze a larger number of samples in one lane. The most advanced method in this regard is the use of unique dual indexing. |
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PacBio Sequel II (Pacific Biosciences)For special applications (for example de-novo genome assembly or when sequencing amplicons and repetitive motifs) longer sequences need to be obtained and SMRT technology used in PacificBiosciences devices can provide this. The length of read can be as much as 50 kb on average with high read quality and we can expect up to approx. 100 Gb data. Compared to other technologies, there is no template amplification step during the preparation of the sample for sequencing and therefore some epigenetic modifications can be recorded directly. A circular DNA molecule serves as a template and DNA polymerase reads it until the sequencing run is finished. Fragments of up to a few tens of kb are therefore repeatedly read and by comparing individual readings performed by the device automatically, the correct identification of every base is achieved. This strategy is called "circular consensus sequencing". By reading 8-10x, reading accuracy is improved to more than 99.99%, which is comparable to that of Illumina. (a picture by PacificBiosciences, adapted)
Note: If you are interested in sequencing by using this technology, please contact us to receive a quote tailored to your project. |
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GridION (OxfordNanopore)A very interesting option for applications requiring long reads are the Oxford Nanopore sequencers. Experiments have now been described in the literature where sequences of up to 2 Mb have been obtained and the read length is essentially limited only by nucleic acid integrity. This technology is based on direct base detection, without the need to amplify the template, hence it has the potential to detect epigenetic modifications directly. OxfordNanopore is still confronted with a reputation for poor quality of sequencing. The truth is that the translation of the signal was and still is a sore point for this technology, since despite constant and rapid progress in this area, reading accuracy is approx. 95%. However, it is still possible to combine long, less accurate sequences with short, accurate ones to obtain far better de-novo assemblies than would provide the short-reads by Illumina themselves. In addition, the progress in signal decoding is very fast and even "old" data can be reanalyzed with new algorithms. |
Sequencing system | Chip/Sequencing chemistry | Sequencing options | Max. length of read | Max. number of reads per chip or flowcell (in millions) | Max number of bases sequenced per chip or flow cell (in Gb) |
NovaSeq6000 (Illumina)1 | SP FlowCell | SE/PE | 2x250 b | 800 | 200 |
S4 FlowCell | 2x150 b | 5000 | 750 | ||
SEQUEL II (Pacific Biosciences)2 | SMRT Cell 8M | HiFi Reads | Q2+ <20 kb | 4 | 60 |
Long Reads | Q2+ >150 kb | 4 | 160 | ||
GridION X5 (Oxford Nanopore Technologies)3 | RevD FlowCell | Native DNA/RNA | ~2 Mb | 3 | 10 |
PCR processed DNA/RNA | ~2 Mb | 3 | 20 |
1 - Sequencing libraries with lower sequence diversity (such as amplicons) face several technological limitations, read here to learn more.
2 - Sequencing output is tightly connected to length of fragments used for library preparation, read here to learn more.
3 - Sequencing output depends strongly on the sample type and sequencing settings, read here to learn more.