Sample Requirements

Our recommendations for your samples

When you plan to ship samples for sequencing to CeGaT, we strongly recommend considering the following sample requirements in order to receive the best results.

Do not hesitate to contact us if you have questions regarding the quantity or the quality of your sample material.

DNA Sequencing

Product

Sample requirements

Quantity

Concentration

Volume

Sequencing
Services

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

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

Microbiome
Analysis

Shotgun Metagenomics

≥250 ng

≥10 ng/µl

≥25 µl

Full Length 16S Sequencing

≥250 ng

≥10 ng/µl

≥25 µl

Translational
Oncology

Comprehensive 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
Services

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
total RNA

≥20 ng/µl

≥50 µl

RIN ≥8

Single-Cell RNA Sequencing

1 million cells

Translational Oncology

CTP FUS Panel

≥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 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 a 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.

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

Cryopreserved cell pellet

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.

Downloads

Sample Requirements & Shipping Guide

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.