DNA Sequencing
Product | Sample requirements | |||
Quantity | Concentration | Volume | ||
Sequencing | Genome Sequencing | ≥500 ng | ≥20 ng/µl | ≥25 µl |
Genome Sequencing (PCR free) | ≥1500 ng | ≥20 ng/µl | ≥75 µl | |
HiFi Genome Sequencing | ≥3000 ng | ≥30 ng/µl | ≥25 µl | |
Exome Sequencing | ≥250 ng | ≥10 ng/µl | ≥25 µl | |
ExomeXtra® Sequencing | ≥250 ng | ≥10 ng/µl | ≥25 µl | |
Panel Sequencing | ≥250 ng | ≥10 ng/µl | ≥25 µl | |
HiFi Panel Sequencing | ≥1000 ng | ≥40 ng/µl | ≥25 µl | |
Epigenomics | Methylation Sequencing | ≥500 ng | ≥20 ng/µl | ≥25 µl |
HiFi Methylation Sequencing | ≥3000 ng | ≥30 ng/µl | ≥25 µl | |
Microbiome | Shotgun Metagenomics | ≥250 ng | ≥10 ng/µl | ≥25 µl |
Full Length 16S Sequencing | ≥250 ng | ≥10 ng/µl | ≥25 µl | |
Translational | Comprehensive Tumor Profiling (CTP) ● CTP WGS ● CTP WES / CTP EXS | ≥500 ng ≥250 ng | ≥20 ng/µl ≥10 ng/µl | ≥25 µl ≥25 µl |
Focused Tumor Profiling | ≥250 ng | ≥10 ng/µl | ≥25 µl | |
Liquid Biopsy | ≥250 ng | ≥10 ng/µl | ≥25 µl | |
TSO500 | ≥500 ng | ≥20 ng/µl | ≥25 µl | |
Immunology | HLA Typing | ≥500 ng | ≥20 ng/µl | ≥25 µl |
Please quantify the concentration of your sample using a fluorometric based method (e.g., Qubit). The quality of your samples should be further assessed by using the Fragment Analyzer or by detecting the optical density. We recommend using samples with high molecular weight DNA that were RNase treated resulting in an OD 260/280 of 1.8 – 2.0. Of course, also fragmented DNA can be sequenced when a sequencing library is prepared according to our specially selected or adapted protocols. All isolated DNA samples should be delivered in molecular biology grade water, 10 mM Tris-HCl pH 8 or Elution Buffer, free of EDTA.
We ask you to send samples in 2 ml screw cap tubes from Sarstedt, or in 1.5 ml Eppendorf tubes. For a sample size ≥ 16 samples, please provide the samples in Eppendorf twin.tec PCR Plates 96, skirted.
Please contact us for further information about submitting lower input amounts or degraded sample material.
RNA Sequencing
Product | Sample requirements | ||||
Quantity | Concentration | Volume | Quality | ||
Sequencing | Coding Transcriptome Sequencing | ≥500 ng | ≥20 ng/µl | ≥25 µl | RIN ≥8 |
Whole Transcriptome Sequencing | ≥500 ng | ≥20 ng/µl | ≥25 µl | RIN ≥3 | |
Small RNA Sequencing | ≥1000 ng | ≥20 ng/µl | ≥50 µl | RIN ≥8 | |
3' Single-Cell RNA Sequencing | 1 million cells | ||||
Single-Cell RNA Sequencing Flex | 2 x 25 µm FFPE scrolls (human) / 1 million fixed cells | ||||
Spatial Transcriptome Sequencing | FFPE tissue block or | ||||
Translational Oncology | Focused Tumor Profiling (FTP) FUS T | ≥50 ng | ≥2.5 ng/µl | ≥20 µl | |
Immunology | T-Cell Receptor Sequencing RNA | ≥250 ng | ≥10 ng/µl | ≥25 µl | |
Single-Cell Immune Profiling | 1 million cells | ||||
Please quantify the concentration of your sample using a fluorometric based method (e.g., Qubit). The quality of your sample should be further assessed by using the Bioanalyzer or by detecting the optical density, resulting in an OD 260/280 ~2.0. RNA samples need to be DNase treated. All RNA samples should be delivered in molecular biology grade water, 10 mM Tris-HCl pH 8 or Elution Buffer, free of EDTA.
We ask you to send samples in 2 ml screw cap tubes from Sarstedt, or in 1.5 ml Eppendorf tubes. For a sample size ≥ 24 samples, please provide the samples in Eppendorf twin.tec PCR Plates 96, skirted.
For 3’ Single-Cell RNA Sequencing and Single-Cell Immune Profiling, we require frozen single-cell suspensions. Each sample should contain 1 million cells in 1 ml cryopreservation medium with cell viability > 90%. No large cell aggregates should be present in the cell suspension prior to cryopreservation. Please provide the samples in Eppendorf twin.tec PCR Plates 96, skirted. For Single-Cell RNA Sequencing Flex, 1 million fixed cells or 2 x 25 µm FFPE scrolls for human samples and 2 x 50 µm FFPE scrolls for murine samples are required prepared according to the preparation guideline.
For Spatial Transcriptome Sequencing, either an FFPE tissue block or FFPE sections on a 10x-approved slide can be provided. The tissue section needs to be placed within the allowable area of the slide and must be provided without a coverslip. Further information about recommended slides and their allowable areas can be found here. If the tissue is larger than 6.5 mm x 6.5 mm, you may mark your area of interest in an H&E image and provide it along with your sample.
Please contact us for further information about submitting lower input amounts or degraded sample material.
Ready to Load Sequencing
Platform | Sample requirements | |
NovaSeq™ X Plus | 120 µl of a 10 nM library pool | |
NovaSeq™ X 6000 | 120 µl of a 10 nM library pool | |
MiSeq | 20 µl of a 10 nM library pool | |
PacBio | 12 µl of a SMRTbell® library | |
The DNA concentration depends on the library insert size: | ||
Library insert size | Concentration | |
≥10 kb | ≥20 ng/µl | |
3000 bp – 9999 bp | ≥6 ng/µl | |
1500 bp – 2999 bp | ≥3 ng/µl | |
500 bp – 1499 bp | ≥1 ng/µl | |
Read to Load Sequencing libraries for the Illumina platforms need to be diluted in 10 mM Tris-HCl, pH 8.5. PacBio libraries need to be delivered in PacBio elution buffer. When we shall use your custom read primers for the sequencing of your library, please send 30 µl of 100 µM concentrated primer.
Samples for DNA and RNA Isolation
Sample type | Sample requirements |
PAXgene blood RNA tubes | 3 – 5 ml |
EDTA/PAX blood tubes | 1 – 2 ml |
Dried blood spot card | 5 spots (100 µl) |
Tissue | 10 – 30 mg (2 mm x 2 mm x 2 mm) |
FFPE-tissue (tumor content > 30%) | 10 sections (10 – 20 µm each) |
FFPE-tissue (tumor content > 30%) for macrodissection | 10 sections (20 – 30 µm each), 1 section 5 µm |
Cell pellet or cells in RLT buffer | 1 million cells |
Saliva | ORAgene DNA tube (DNA Genotek) |
Buccal swab | ORAcollect DNA tube (DNA Genotek) |
Fecal samples | CeGaT’s sampling kit |
Stool samples must either be placed fresh in DNA/RNA shield according to our sampling kit (then no freezing necessary) or frozen directly. Even shorter storage periods under different conditions can significantly falsify the results.
General Sample Shipment Recommendations
When sending your samples for sequencing, please consider the following shipment conditions:
- DNA samples should be shipped chilled at 4° C or frozen.
- RNA samples should be shipped frozen on dry ice.
- Sequencing libraries should be shipped frozen on dry ice.
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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.
