The introduction of next-generation sequencing (NGS) fundamentally changed how we approach scientific and diagnostic issues. Next-generation sequencing’s potential to produce large amounts of sequence data for a relatively low price led to a series of groundbreaking discoveries, ushering the field of genomics into a new era. With the development of further technologies, such as single-molecule real-time (SMRT) sequencing, part of third-generation sequencing (TGS), there is now no obstacle to elucidating large structural variants.
Five Illumina NovaSeq™ 6000 and four NovaSeq™ X Plus are available to us for your high-throughput short-read sequencing. Furthermore, we can provide sequencing for smaller projects on two MiSeq Sequencers. Regarding high-quality long-read sequencing, we have established the PacBio platforms Revio and Sequel® IIe. Taken together, we can offer a comprehensive range of sequencing applications for research projects and pharmaceutical studies.
CeGaT has specialized in the use of NGS since its foundation. We continually invest in the latest technologies to aim for the most accurate and reliable results for our customers. Our profound diagnostic knowledge and bioinformatic expertise complement the state-of-the-art lab platforms and sequencing technology. We offer a unique combination of know-how, the latest technology, and premium service for your projects.
Our team has substantial experience with human and animal samples, as well as a wide range of expertise in processing plant, fungi, and microbial samples.
Based on many years of experience working with NGS technology, we have developed significant expertise in high-throughput processing for thousands of samples. Our laboratories are equipped with liquid-handling workstation platforms from Hamilton. These platforms can be individually programmed, allowing specific adaptations of our protocols based on your requirements.
The multiplexing strategy established in our laboratory enables the sequencing of hundreds of samples in a single sequencing run, or even in a single lane, and thus represents true high-throughput sequencing.
We would be happy to demonstrate the feasibility of your high-throughput projects in a pilot study.
Illumina Sequencing Platforms
Illumina NovaSeq™ X Plus System
- For high-throughput sequencing with exceptional accuracy
- Ideal to perform larger studies and more data-intensive projects
- Produces up to 16 Tb of data in 48 hours per dual flow cell run
Find a performance comparison of our NovaSeq™ devices here.
Used under license from Illumina, Inc. All rights reserved.
Illumina NovaSeq™ 6000 System
- For high-throughput sequencing with the lowest cost-per-base
- Ideal for large-scale and deep-sequencing projects
- Produces up to 6 Tb of data in 20 hours
Find a performance comparison of our NovaSeq™ devices here.
Used under license from Illumina, Inc. All rights reserved.
Illumina MiSeq™ System
- For smaller sequencing projects with multiple configurations
- Read length up to 2 x 300 bp
Used under license from Illumina, Inc. All rights reserved.
Illumina Workflow
Preparation of a DNA Library
For DNA library preparation, DNA is first fragmented and then undergoes end-repair. Next, adapters are ligated to both ends of the repaired DNA fragments, completing the DNA library preparation process. These adapters contain specific sequence motifs that are essential for the subsequent steps, which include clonal amplification and actual sequencing.
Used under license from Illumina, Inc. All rights reserved.
Clonal Amplification
Illumina sequencing technology utilizes the so-called bridge-PCR for clonal amplification of individual DNA fragments. DNA fragments are linked via the adapters to the flow cell, and then the bridge-PCR is carried out. Oligos bound to the flow cell function as primers, and each individual DNA fragment forms a separate cluster of identical DNA fragments.
Used under license from Illumina, Inc. All rights reserved.
Sequencing
Illumina’s Sequencing by Synthesis (SBS) method utilizes the Cyclic Reversible Termination (CRT) approach. In this process, each of the four nucleotides is attached to a different dye and modified with a terminator group. During each reaction cycle, all four nucleotides are presented simultaneously to the polymerase for strand synthesis. Once a complementary nucleotide is incorporated, elongation stops due to the blocking effect of the terminator group. The dyes associated with each nucleotide can then be detected using imaging techniques. The dye and the terminator group are cleaved, allowing a new synthesis cycle to begin. This process enables the simultaneous determination of the sequence for each cluster, base by base.
Used under license from Illumina, Inc. All rights reserved.
PacBio Platforms
PacBio Revio System
- For high-quality long-read HiFi sequencing
- Output reaches up to 480 Gb and 99.95% read accuracy
- Methylation calling based on kinetics
Image courtesy of PacBio
PacBio Sequel® IIe System
- For high-quality long-read HiFi sequencing
- Sequencing projects with reads up to 25 kb and 99.9% accuracy
PacBio Workflow
Preparation of Sequencing Library
Hairpin sequencing adapters are ligated to high-molecular-weight, double-stranded DNA, creating so-called circularized SMRTbells®. Subsequently, primers and the polymerase are bound to the adapter sequence. The successfully prepared library is now loaded on the SMRT® cell.
Image courtesy of PacBio
Sequencing
The SMRT® cell contains millions of wells called zero-mode waveguides (ZMWs). Single DNA molecules are immobilized in these wells, and the polymerase begins to incorporate fluorescently labeled nucleotides around the cycle. In this process, light is emitted, and nucleotide incorporation is measured in real-time. When sequencing in circular consensus mode (CCS), highly accurate long reads (HiFi) are generated because SMRT sequencing can sequence the same DNA molecule multiple times.
Image courtesy of PacBio
10x Genomics® Platform
Our 10x Genomics® platforms are not used for sequencing itself; that occurs on our other platforms. Instead, they are designed for sample preparation for sequencing at the single-cell level.
10x Genomics® Chromium™ X Controller
- For analysis of single cells at a massive scale
- Microfluidics system to capture tens of thousands of single cells per sample
Image courtesy of 10x Genomics, Inc.
10x Genomics® Chromium Workflow
Generation of Gel Beads in Emulsion
On the chromium chip, gel beds in emulsions (GEMs) are generated by combining 10x barcoded gel beads, a MasterMix containing cells, and partitioning oil, leading to the formation of droplets containing one 10x barcoded bead and one cell (Single Cell GEMs).
Image courtesy of 10x Genomics, Inc.
Preperation of a Library and Sequencing
After GEM generation, the gel beads dissolve, releasing oligos and enzymes into the solution, which enables amplification and results in 10x barcoded cDNA within each droplet.
After barcoding, the barcoded fragments are pooled together, and the final library construction is performed in bulk. Sequencing of the libraries is conducted on one of our Illumina sequencing systems.
Image courtesy of 10x Genomics, Inc.
10x Genomics® Visium Cyt Assist
- For high-resolution spatial gene expression
- Combines transcription analysis with tissue morphology
Image courtesy of 10x Genomics, Inc.
10x Genomics® Visium Workflow
Tissue Preparation
Ultrathin tissue slices from FFPE tissue samples are transferred to microscopic glass slides, which are stained to visualize the spatial tissue morphology.
Probe Transfer using the Visium CytAssist
A probe mix is dispensed onto the slide, and the tissue RNA is ligated to the probes. Using the Visium CytAssist, a brightfield image is taken, followed by the transfer of the transcriptomic probes to the capture area of a Visium Slide. The probes are extended by localization encoding barcodes and subsequently released.
Library Preparation and Sequencing
The barcoded probes are used for library preparation and sequencing with one of our Illumina platforms. Finally, the gene expression information is superimposed with the histological image, resulting in a gene expression map.
Image courtesy of 10x Genomics, Inc.
Contact Us
Do you have a question or are you interested in our service? Feel free to contact us. We will take care of your request as soon as possible.
Start Your Project with Us
We are happy to discuss sequencing options and to find a solution specifically tailored to your clinical study or research project.
When getting in contact, please specify sample information including starting material, number of samples, preferred library preparation option, preferred sequencing depth and required bioinformatic analysis level, if possible.
