Next Generation Services
Next-generation sequencing (NGS), also called massively parallel or deep sequencing, stands for a DNA/RNA sequencing technology that is much quicker and cheaper than the previously used Sanger sequencing and has revolutionized genomic research. A good summary for users new to NGS technologies can be found in the document "An Introduction to next generation technology". Please see below for an overview of our services and how to book them.
Next-generation sequencing (NGS), also called massively parallel or deep sequencing, stands for a DNA/RNA sequencing technology that is much quicker and cheaper than the previously used Sanger sequencing and has revolutionized genomic research. A good summary for users new to NGS technologies can be found in the document "An Introduction to next generation technology". Please see below for an overview of our services and how to book them.
Service Overview
Our NGS platform provides sequencing services for both DNA and RNA analysis as follows:
- Automated DNA/RNA Isolation
- Automated library preparation for WGS, Exome-Seq, and RNA-Seq (mRNA and total RNA)
- NGS Sequencing full service
- Sequencing only service (users provide Illumina kit)
- Single Cell Sequencing
- Long read Sequencing (in development)
- Method development
- Consultation on experimental design and troubleshooting
- Support for the submission of grants and publications
- Access to self-service instruments (trainied users only)
To start a new project we require users to book an appointment for a consultation session. Please send us the project proposal form in advance to this meeting. After agreeing the best way forward for the project, the core will provide a quote and estimated time line for completion. Details about the workflow are summarized in our NGS guidelines.
Users can furthermore access some of our instruments for self-service use after prior training by facility staff. Please contact us to arrange training.
NGS services
The Illumina NovaSeq 6000 sequencing system can run four different flow cell types (SP, S1, S2, and S4) and various read length combinations to enable effective throughput as needed. Flow cell features are summarized in tables 1 and 2 below. For additional flexibility, the NovaSeq Xp workflow supports individual lane loading for sequencing different libraries in each flow cell lane. The core will determine the best solution with each customer.
Table 1: Flowcell-Options for the Illumina NovaSeq 6000
Flow Cell | SP | 1 | S2 | S4 |
Available sizes (In cycles) | 100 200 300 500 | 100 200 300 | 100 200 300 | 35 200 300 |
Single or Paired-reads | 700-800 M | 1.3 – 1.6 B | 3.3 – 4.1 B | 8 – 10 B |
Number of lanes | 2 | 2 | 2 | 4 |
Please note: The NovaSeq 6000 v1.5 Kit contains reagents for 38 additional cycels to support the use of Unique Dual Indexes (UDIs) and Unique Molecular Identifiers (UMIs).
Table 2: Estimated sample throughput for frequently used applications
Flow Cell Type | SP | S1 | S2 | S4 |
Human Genomes per Run | ~2 | ~4 | ~10 | ~24 |
Exomes per Run | ~20 | ~40 | ~100 | ~250 |
Transcriptomes per Run | ~16 | ~32 | ~82 | ~200 |
Source: https://emea.illumina.com/systems/sequencing-platforms/novaseq/specifications.html
We offer the options below for sequencing on the NovaSeq, routinely the core will order all reagents and invoice for the complete service:
- Sequencing plus library-Prep from isolated DNA/RNA
Book using forms: DNA/RNA Seq Order und DNA/RNA Sample list - Sequencing of customer-made libraries
Book using forms: Library Seq Order und Library-List - Sequencing only service (Customer delivers Illumina Sequencing Kits and ready made library)
Book using forms: Library Seq Order und Library-List
The Illumina NextSeq 1000 sequencing system can run two different flow cell types (P1 and P2) and various read length combinations to enable effective throughput as needed (100 Mio reads in P1 and 400 Mio reads in P2). This benchtop sequencer creates the perfect output bridge between the low throughput Miseq sequencer (up to 25Mio reads) and the high throughput NovaSeq 6000 (starting with 800 Mio reads). Flow cell features are summarized in tables 1 and 2 below.
Table 1: Sequencing Output Per Flow CellNextSeq 1000
Flowcell type | P1 | P2 |
2 x 50 | N/A | 40 Gb / 400 Mio reads |
2 x 100 | N/A | 80 Gb / 400 Mio reads |
2 x 150 | 30 Gb / 100 Mio reads | 120 Gb / 400 Mio reads |
Table 2: Estimated Sample Throughput for Key Applications
Flow Cell Type | P1 | P2 |
Small Whole-Genome Sequencing (300 cycles) | ~7 | ~30 |
Whole-Exome Sequencing (200 cycles) | ~4 | ~16 |
Single-Cell RNA-Seq (100 cycles) |
| ~2-10 |
Source: https://emea.illumina.com/systems/sequencing-platforms/nextseq-1000-2000/specifications.html
We offer the services below for sequencing on the Nextseq 1000, routinely the core will order all reagents and invoice for the complete service:
- Sequencing plus library-Prep from isolated DNA/RNA
Book using forms: DNA/RNA Seq Order und DNA/RNA Sample list
- Sequencing of customer-made libraries
Book using forms: Library Seq Order und Library-List
- Sequencing only service (Customer delivers Illumina Sequencing Kits and ready made library)
Book using forms: Library Seq Order und Library-List
The Illumina MiSeq is a benchtop sequencer ideal for projects requiring a limited number of reads (up to 25 M). It is also frequently used for prokaryotic sequencing projects. User can order runs as a full service or operate the Core’s MiSeq independently after introductory training. Please contact us prior to first use. The MiSeq's flow cell has only one lane, for technical details see tables 1 and 2 below (Source: https://emea.illumina.com/systems/sequencing-platforms/miseq/specifications.html)
Service booking:
Please complete forms "MiSeq Library Seq Order" and "Library list" and email these to the core.
Table 1: Miseq Cluster generation and Sequencing
Table 2: Reads passing filter
For high throughput, standardized library preparation we use two Agilent Bravo NGS workstations (pre and post PCR) to process up to 96 samples per run.
Our library prep portfolio:
- Whole genome and whole exome library preparation for DNA (Illumina DNA PCR-Free Prep
- Tagmentation library,
- Agilent V6 Exome and Twist exome/panels
- mRNA and total-RNA library preparation (TruSeq Stranded mRNA and TruSeq Stranded Total RNA as well as Illumina Stranded mRNA prep Ligation).
Crucially important for successful and high quality library prep is the exact mesurement of the concentration and the quality of the submitted DNA/RNA. Input DNA and RNA quantities specified in table 1 apply only if samples were quantified by a fluorometric method (e.g. Qubit, PicoGreen, RiboGreen). If a spectrophotometer (e.g. Nanodrop) was used, we suggest submitting twice the requested amount of sample since this type of measurement is often unreliable. In any case, sample amounts higher than the minimum requirements will improve the library complexity and thus the quality of the data.
Prpared libraries are ready to be sequenced on the Miseq or Novaseq 6000 depending on the desired output. If you have a large project that would require a different library preparation method, get in contact with us to discuss the automation possibilities.
Booking services:
Please complete the forms "Library order form" und "DNA/RNA sample list" and email these to the core.
Table 1: Minimum amount and quality of nuclic acids for various library prep protocols
Library Type | Quality | Quantity* | Volume* | Buffer |
DNA PCR-Free Prep, Tagmentation library | double-stranded non-degraded A260/280 ratio > 1.8 RNA-free | 500 ng | In 25 µl | EB, H2O |
Agilent V6 Exome | double-stranded non-degraded A260/280 ratio > 1.8 RNA-free | 3µg | In 130 µl | EB, H2O |
Twist Exome/Panels | double-stranded non-degraded A260/280 ratio > 1.8 RNA-free | 50 ng | In 10 µl | EB, H2O |
Stranded mRNA (Truseq) | RIN > 7 DNA-free | 300 ng- 1µg | In 50 µl | non-DEPC treated H2O |
Stranded Total RNA (Truseq) | RIN > 7 DNA-free | 300 ng- 1µg | In 10 µl | non-DEPC treated H2O |
Stranded mRNA (ligation) | RIN > 7 DNA-free | 300 ng- 1µg | In 25 µl | non-DEPC treated H2O |
Our Equipment
Instruments run by Core Staff only:
- Illumina NovaSeq 6000
- Illumina Nextseq 1000
- 10x Genomics Chromium
- Agilent Bravo robotic workstations (library prep)
- TECAN Fluent robotic workstation
- Perkin Elmer LabChip GX Touch HT
- QC-Instruments (Qubit, Nanodrop, Covaris200)
User accessible instruments
- Illumina MiSeq Sequenzierer
- Agilent Bioanalyzer 2100 (Cartridges available upon request at extra costs)
