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QIAGEN Sequencing Services

For the highest standards in contract DNA sequencing
  • Single-read services
  • Verification services
  • De novo sequencing
  • Genome services
  • Special services
QIAGEN offers a wide range of sequencing services. Our single-read services enable routine sequencing of plasmids, PCR products, and expressed sequence tags (ESTs).  QIAGEN's verification services allow resequencing and comparison with reference sequences, while our de novo sequencing services enable analysis of unknown sequences, from single stranded primer walking to patent quality. Our genome services are suitable for custom/tailored projects abd allow access to genomic sequence information. Additonal special services include siRNA hairpin runs, genomic DNA sequencing, mutation detection, DNA purification, bioinformatics, and whole genome amplification.

 

Cat No./ID: SeqService
QIAGEN Sequencing Services
Contract DNA sequencing services: Single-Read Services, De Novo Sequencing, Verification Services, Genome Services, Special Services
Performance
QIAGEN delivers the highest standard in contract DNA sequencing.

QIAGEN Sequencing Services — Single-Read Services
Single-read sequencing services (96-well format) are ideal for small-scale analysis of cDNAs and ESTs, identification of clones, and verification of vector/insert transitions. All single read services include extensive documentation of the sequencing data, which is available via e-mail, FTP file, or on CD.

Easy-read sequencing
Easy-read sequencing is the service of choice for standard sequencing reactions. This highly automated service allows rapid processing of purified DNA samples. Easy-read sequencing is offered as tube service, or as a plate service for sequencing of samples in batches of 96. Easy-read sequencing does not include template purification (i.e., purification of PCR fragments or plasmid preparations prior to sequencing reaction setup). Sequencing reactions are set up using the DNA concentration information provided by the customer. All sequencing results are subjected to quality control using Phred20 algorithms (typically 600–900 bases of Phred20 controlled sequence are obtained). Quality documentation including raw data and quality clipped sequence data is delivered via e-mail, ftp, or on CD. Failed sequences are not repeated. This service provides high-quality single-stranded sequencing data 1–2 working days after receipt of purified DNA.

Easy-sead sequencing provides:
• Single sequencing reaction
• Data collection to maximum read length
• Typical read length up to 600–900 nt
• Phred20 quality
• >99% accuracy
• Without template purification
• Without incoming DNA quality check
• No repeat reaction
• Documentation of sequencing results

Single-read sequencing
If your sequencing reaction needs special attention we recommend our single-read sequencing Service. The sequencing results are manually checked and any failed sequencing reactions are repeated using modified sequencing parameters. This means that you get the most reliable service even from difficult samples (e.g., samples with high GC content). This service is offered with or without template purification as a plate service for sequencing of samples in batches of 96. Single-read sequencing with template preparation includes the purification of PCR fragments or plasmids prior the sequencing reaction setup. Templates are quality controlled to ensure an optimal sequencing reaction setup. All sequencing results are subjected to quality control using Phred20 algorithms (typically 600–900 bases of Phred20 controlled sequence are obtained). Quality documentation including raw data and quality clipped sequence data is delivered via e-mail, ftp, or on CD. Failed sequences are inspected by our experienced team and repeated using modified sequencing parameters. This service provides high-quality single-stranded sequencing data in 2–3 working days (3–4 working days with template purification).

Single-read sequencing provides:
• Single sequencing reaction
• Incoming DNA quality check
• Repeat reaction, if necessary
• Data collection to maximum read length
• Read length up to 600–900 nt
• Phred20 quality
• >99% accuracy
• Documentation of sequencing results

BAC end-sequencing
End-sequencing of BACs, PACs, or cosmids. The results are manually checked by our experienced team and failed sequencing reactions are repeated using modified sequencing parameters. This means that you get the most reliable service even from difficult samples (e.g., samples with high GC content). This service is offered with or without template purification as a plate service for sequencing of samples in batches of 96. The delivery time is approximately 2–3 working days (3–4 working days with template purification). 

BAC End-Sequencing provides:
• Single sequencing reaction
• Incoming DNA quality check
• Repeat reaction if necessary
• Data collection to maximum read length
• Read length up to 600–900 nt
• Phred20 quality
• >99% accuracy
• Documentation of sequencing results

siRNA hairpin run
siRNA expression vector templates exhibit secondary structures that are often difficult to sequence. QIAGEN has developed a special sequencing reaction that enables accurate sequencing of siRNA expression vector templates. The results are manually checked by our experienced team and failed sequencing reactions are repeated using modified sequencing parameters. This service is offered with or without template purification as a plate service for sequencing of samples in batches of 96. The delivery time is approximately 2–3 working days (3–4 working days with template purification). 

siRNA hairpin run provides:
• Single sequencing reaction
• Incoming DNA quality check
• Repeat reaction if necessary
• Data collection to maximum read length
• Read length up to 400-600 nt
• Phred20 quality
• >99% accuracy
• Documentation of sequencing results

Available sequencing services
Feature Easy-Read Sequencing  Single-Read Sequencing  BAC End-Sequencing siRNA Hairpin Run
Sequencing reaction X X X X
Template purification   Optional Optional Optional
Template quality check   X X X
Repeat reaction   X X X
Data collection to maximum read length X X X X
Read length (nt) 600–900 600–900 600–900 400–600
Quality Phred20 Phred20 Phred20 Phred20
Accuracy >99% >99% >99% >99%
Documentation E-mail, FTP, or CD >99% E-mail, FTP, or CD E-mail, FTP, or CD
Turn around time (working days) 1 2 (3–4 with template purification) 2 (3–4 with template purification) 2 (3–4 with template purification)

QIAGEN Sequencing Services — Verification Services

Verification sequencing
Verification sequencing provides confirmation of an already known DNA sequence, as well as detection of mutations or deletions. Both DNA strands are completely sequenced using custom primers based on a provided DNA sequence. The sequence is verified against the provided reference sequence.

Verification sequencing provides:

  • Template purification
  • Design and synthesis of internal primers
  • Complete sequencing of both DNA strands (>99.995% accuracy)
  • Sequence editing and assembly
  • Verification of the sequence against a customer-provided reference sequence
  • Sequence data in publication-ready format
  • Project documentation
  • Electronic data delivery
  • Experienced and reliable support
Alternatively, BACs, PACs, or cosmids can be sequenced to a predetermined sequence coverage including assembly. Please ask for this service.

Verification sequencing Under GLP-like conditions
Verification sequencing Under GLP-like conditions provides confirmation of an already-known DNA sequence including comprehensive documentation of the sequencing results. Both DNA strands are completely sequenced to at least 4-fold coverage. The sequence is verified against a provided reference sequence. Documentation can be used for patent applications, litigations, or FDA submissions.

GLP-like verification sequencing comprises:

  • Template purification
  • Design and synthesis of internal primers
  • Complete sequencing of both DNA strands (>99.995% accuracy)
  • 4-fold coverage
  • Sequence editing and assembly
  • Verification of the sequence against a customer-provided reference sequence
  • Sequence data in publication-ready format
  • Comprehensive project documentation
  • Electronic data delivery
  • Experienced and reliable support

QIAGEN Sequencing Services — De Novo Sequencing

QIAGEN offers a number of de novo Sequencing services, providing accurate sequence data from unknown DNA. Sequencing quality ranges from single stranded primer walking for preliminary analysis to complete sequence analysis for patent submission.

De novo publication-ready sequencing
The de novo publication-ready sequencing service provides publication-ready DNA sequence from unknown template DNA. Both DNA strands are completely sequenced and the sequence is fully edited and assembled. Depending on the project size, different sequencing strategies can be applied (i.e., primer walking for plasmids or shotgun sequencing cosmids, BACs, or PACs).

If you provide us with an E. coli host strain harbouring the construct of interest, we will apply the best strategy to determine the sequence and we will send back the sequencing results within 20 working days.

De novo publication-ready sequencing provides:

  • Complete sequencing of both DNA strands (>99.995% accuracy)
  • Template purification
  • Design and synthesis of internal primers or shotgun library generation
  • Sequence editing and assembly
  • Sequence data in a publication-ready format
  • Applicable project documentation
  • Electronic data delivery
  • Experienced and reliable support
Alternatively, BACs, PACs, or cosmids can be sequenced to a predetermined sequence coverage including assembly. Please ask for this service.

De novo investigation-grade sequencing
De novo investigation-grade sequencing provides a single stranded primer walking approach for preliminary analysis of unknown sequences. The target DNA is sequenced to single-strand coverage and the sequence is edited and assembled. We guarantee a sequence quality score of Phred20 (i.e., an error rate of 1:100 nucleotides) for all assigned bases.

De novo investigation-grade sequencing provides:

  • Complete sequencing of one DNA strand (>99% accuracy)
  • Template purification
  • Design and synthesis of internal primers
  • Sequence editing and assembly
  • Sequence data in a publication-ready format
  • Applicable project documentation
  • Electronic data delivery
  • Experienced and reliable support

De novo publication sequencing under GLP-like conditions

De novo publication sequencing under GLP-like conditions provides double stranded sequencing including comprehensive documentation. Both DNA strands are completely sequenced to at least four fold coverage. Documentation can be used for patent applications, litigations, or FDA submissions.

If you provide us with an E. coli host strain harbouring the construct of interest, we will apply the best strategy to determine the sequence and we will send back the sequencing results within 20 working days.

De novo publication sequencing Under GLP-like conditions provides:

  • Complete sequencing of both DNA strands (>99.995% accuracy)
  • At least 4-fold sequencing coverage
  • Template purification
  • Design and synthesis of internal primers or shotgun library generation
  • Sequence editing and assembly
  • Sequence data in a publication-ready format
  • Extensive project documentation report
  • Electronic data delivery
  • Experienced and reliable support


QIAGEN Sequencing Services — Genome Services


QIAGEN’s genome service provides large-scale genomic DNA sequencing including the sequencing of whole genomes.

The genome aervice provides:
  • De novo DNA sequencing of complete microbial genomes
  • Draft sequencing of genomes to a defined coverage
  • Genomic DNA sequence comparisons between related bacterial strains
QIAGEN has participated in various genome projects. Our genome-sequencing service includes the production of BAC or shotgun-cloned libraries or whole genome shotgun libraries, high-throughput sequencing, and data annotation. 

QIAGEN sequencing services has become a well-established participant in many publicly funded and private genome projects. The following table shows our contribution to selected projects:

Project overview
Project (Database links)
References
Saccharomyces cerevisiae (European Yeast Genome Project) Bussey, H. et al. (1997) Nature 387 (suppl.), 103.
Dujon, B. et al. (1994) Nature 369, 371.
Dujon, B. et al. (1997) Nature 387 (suppl.), 98.
Johnston, M. et al. (1997) Nature 387 (suppl.), 87.
Philippsen, P. et al. (1997) Nature 387 (suppl.), 93.
Bacillus subtilis Kunst, F. et al. (1997) Nature 390, 249.
Schizosaccharomyces pombe Wood, V. et al. (2002) Nature 415, 871
Arabidopsis thaliana Bevan, M. et al. (1998) Nature 391, 485.
The EU Arabidopsis Genome Sequencing Consortium et al. (2000), Sequence and analysis of chromosome 4 of the plant Arabidopsis thaliana, Nature 402, 769
Leishmania major Ivens, A.C. et al. (2005) Science 309, 436
Full Length Human cDNA Project Wiemann, S et al. (2001) Genome Research 11, 422
Pseudomonas putida (joint effort with TIGR, MHH, DKFZ, GBF; genome size: 6.2 Mbps) TIGR Microbial Database


Other large-scale genome-sequencing projects
Project (Database links)
Strategy Funding
Alcanivorax borkumensis Whole genome shotgun Univ. Bielefeld
Aspergillus niger BAC-by-BAC DSM Food Specialties
Bacillus clausii Whole genome shotgun Genencor
Chlamydia pneumoniae Whole genome shotgun Byk-Gulden
Flavobacterium psychrophilum Whole genome shotgun
Geobacillus thermoleouorans Whole genome shotgun
Hansenula polymorpha BAC-by-BAC Rhein Biotech
Listeria seeligeri Whole genome shotgun
Propionibacterium freudenreichii Whole genome shotgun DSM + FCDF
Sorangium cellulosum Whole genome shotgun
Xanthomonas campestris Whole genome shotgun

One critical factor for successful genome sequencing is the generation of whole genome shotgun libraries. Our shotgun libraries are of an extremely high quality. Shotgun libraries with different insert sizes are used to establish the sequence either up to publication quality (>99.995% sequence accuracy) or up to a certain genomic coverage.

We also offer various bioinformatic analyses including complete annotation and comparative sequencing.

For large-scale projects, a dedicated project management plan and team is instigated to carry out all necessary work in due speed and to the highest accuracy and quality levels.
 
QIAGEN Sequencing Services — Special Services

QIAGEN offers a number of special services that complement our sequencing service. These services include the sequencing of templates with a high degree of secondary structure, the sequencing of genomic DNA, the detection of mutations, DNA purification, whole genome amplification, and bioinformatic analysis.

siRNA hairpin run
siRNA expression vector templates exhibit secondary structures that are often difficult to sequence. QIAGEN has developed a special sequencing reaction that enables accurate sequencing of siRNA expression vector templates. The results are manually checked by our experienced team and any failed sequencing reactions are repeated using modified sequencing parameters. This service is offered with or without template purification as a plate service for sequencing of samples in batches of 96. The delivery time is approximately 2–3 working days (3–4 working days with template purification).


siRNA hairpin run provides:

  • Single sequencing reaction
  • Incoming DNA quality check
  • Repeat reaction (if necessary)
  • Data collection to maximum read length
  • Read length up to 400-600 nt
  • Phred20 quality
  • >99% accuracy
  • Documentation of sequencing results

Genomic DNA Sequencing
Primer walking directly on genomic DNA provides a means of efficiently closing gaps between contigs. We use special reaction parameters to directly sequence prokaryotic or viral DNA with genome sizes <5 mb.

Genomic DNA sequencing provides:

  • Sequencing reaction directly on prokaryotic or viral DNA (<5 mb)
  • Single sequencing reaction with special sequencing parameters
  • Quality check of incoming DNA
  • Repeat reaction (if necessary)
  • Data collection to maximum read length
  • Typical read lengths of 200-700 nt depending on genome size and DNA quality
  • Phred20 quality
  • >99% accuracy
  • Documentation of sequencing results

Mutation detection service
Sequencing remains the method of choice for mutation detection and profiling. QIAGEN Sequencing Services offer a Mutation detection service tailored to your specific needs. Our bioinformatics experts will perform all steps including assay design and primer synthesis, PCR optimization and amplification, DNA purification, bi-directional DNA sequencing, quality scoring, automated mutation calling, and sequence analysis.


The mutation detection service provides:

  • Quality check of incoming DNA
  • Assay development and validation
  • Primer design
  • Optimization of PCR amplification conditions
  • PCR amplification and product purification
  • Bi-directional DNA sequencing reaction
  • Comparison with reference sample
  • Quality report and mutation report

DNA purification service
Don't waste your time with routine work. QIAGEN offers both small- and large-scale DNA preparation services from a variety of different starting materials.


The DNA purification service provides:

  • Plasmid DNA preparation from bacterial cultures
  • Cosmid, fosmid, BAC preps
  • PCR purification
  • DNA purification from EDTA-blood
  • DNA purification from buccal swabs
  • Preparation in 96- or 384-well plate format

Whole genome amplification service
If you do not have enough starting material for your downstream analysis, we recommend amplification of your genomic DNA. The REPLI-g service — based on proven REPLI-g whole genome amplification technology — allows the amplification of unlimited amounts of DNA from limited samples with minimal sequence bias. A stringent quality control assay provides information on the quality of the amplified DNA, enabling reliable predictions for the success of your downstream assay to be made. For more information, or to find out how to take advantage of this service click here.


Bioinformatic analysis
Every sequencing project has special requirements for bioinformatic resources (e.g., large scale BLAST analysis, EST clustering, custom primer design, or annotation). The QIAGEN Sequencing Service provides in-depth bioinformatic analysis of DNA sequences for sequencing projects of any scale
Principle
Simply send your sample and take advantage of QIAGEN's expertise. From sample receipt, DNA purification and QC of purified DNA to sequencing and data analysis and delivery, QIAGEN offers a one-stop service for all your contract manufacturing and sequencing needs.
Procedure
To get started, obtain a quotation for your project request. If you are a new customer, please contact QIAGEN Sequencing Services (QSS) for pricing information.


Complete an order request
Please download our order form in Excel format. Order forms in excel format should be e-mailed to sequencing@qiagen.com in advance. A hardcopy MUST BE submitted with every package shipped to QIAGEN.

Sample submission and shipping
Please provide the following DNA concentrations per sequencing reaction:

  • Plasmid (3-10 kb): 0.5 µg
  • Plasmid (10-20 kb): 1 µg
  • Cosmid (30-45 kb): 3 µg
  • BAC/ PAC: 3 µg
  • Bacterial genomic DNA: 10 µg
  • PCR product (< 500 bp): 0.1 µg
  • PCR product (500-2000 bp): 0.2 µg
  • PCR product (>2000 bp): 0.5 µg
  • Unpurified PCR product: send the whole PCR reaction (50-100 µl)
If you submit your own primers, we need 20 pmol primer per sequencing reaction. Please supply your primer dissolved in water (5 pmol/µl),

Procedures for shipping clones and DNA from countries outside the EU to QIAGEN Sequencing Services
QIAGEN has selected FedEx as our preferred carrier to Germany.

Please follow this checklist carefully to ensure that your samples are processed efficiently through FedEx and German customs.

A padded shipping envelope may be used for small numbers of tubes or plates. For larger shipments, boxes may be used but must accommodate the FedEx International Waybill and Commercial Invoice.

  • Securely pack samples for shipment with padding to prevent damage to vials or culture plates. Tighten caps on liquid samples and seal with Parafilm to prevent leakage. No samples with a biohazard rating above the lowest level (i.e., BL1, S1, P1, L1) should be shipped. Clones from non-infectious organisms in standard laboratory strains of E. coli are all acceptable.
  • On a FedEx International Air Waybill, write in the date of shipment, your account number, and your complete address and phone number in box 1.
  • In box 2, enter: André Bahr
  • Phone +-49-2103-29-16345
  • QIAGEN Genomic Services / Sequencing
  • QIAGEN GmbH
  • Max-Volmer-Str. 4
  • Hilden. Germany D-40724
  • Enter a description and harmonized code for the contents of your shipment in box 3. For bacterial cultures, enter “3002.90.50 Microorganisms, E. coli cultures, non-infectious, non-hazardous, for research purposes only, contained in (give number) vials/plastic plates.” For DNA samples (e.g. genomic DNA or PCR products), write “2933.39.90 Nucleic Acids, contained in (give number) vials/plastic plates.” The value of the shipment stated for customs should not total more than $25 per shipment. (This is basically the value of the plastics; the clones themselves, although of course worth a lot to you, are considered by customs as having no value. The $25 or less value makes it easier to get the shipment through customs, as there will be no duties applied.) The box marked “No SED required, value $2500 or less” should be checked.
  • In box 4, select the shipping option that works best for you. Please use FedEx First or FedEx Priority for shipments containing dry or wet ice.
  • In box 5, indicate your packaging option. No special handling should be indicated in box 6 if package is shipped early in the week. We recommend that all shipments containing dry or wet ice be shipped on either Monday or Tuesday.
  • Indicate your preferred payment option in box 7. (Note: QIAGEN does not routinely pay for sample shipping.)
  • For tracking purposes, please enter your Purchase Order Number in box 8 under “Your Internal Billing Reference”.
  • Enter your signature in box 9. Check that the shipment does not contain dangerous goods (Note: QIAGEN cannot accept materials from pathogenic organisms). Shipments containing dry ice will require a FedEx dry ice label.
  • On a Commercial Invoice form, enter the information requested to match the information on the FedEx International Waybill. In particular, enter the exact same description used in box 4 under "Description of Goods". Under “No. Units” write the number of plates or vials, under "Unit Value", write the value of each plate or vial, and provide the multiplied value in the total value column.
  • Under “Terms of Sale (Incoterm)”, write “CIF”. This indicates that the basis of the “sale” (any international transfer) is cost, insurance, and freight. Fill in the number of packages, weight, shipping cost, and insurance ($0).
  • Sign your name under "Shipper’s Signature".
  • Insert the International Waybill and two copies of the Commercial Invoice in a plastic adhesive FedEx envelope and affix it to your shipping envelope or other container. Call FedEx Customer Service (1-800-Go-FedEx) for a pick up or your nearest drop off location. For further shipping information, contact FedEx International Services (800-247-4747).
Phone contact at QIAGEN Genomics in Germany (also for contacts by carrier or customs):
Andre Bahr: +49-2103-29-16345
Anke Henne: +49-2103-29-16234


Retrieval of results
DNA sequencing results are made available via e-mail or secure (password protected) web download, typically 24-48 hours after samples have arrived in our labs.


Contact/shipping information
Please send your samples to:

QIAGEN Genomic Services
Sequencing Services
QIAGEN Str. 1
D-40724 Hilden
Germany

IMPORTANT: Do not forget to indicate the sender when you send your samples! This information is essential so that we can correctly match your samples to your order form. Alternatively, please include a hard copy of the order form when you send your samples to us.
You will receive an acknowledgement of your order by e-mail when we receive your samples.
If you have any questions, please do not hesitate to call your local Technical Service or the Genomic Services Department in Germany at +49 2103 29 16234 or +49 2103 29 16345.
Applications

QIAGEN Sequencing Services can be used for:

  • Routine sequencing of plasmids, PCR products, expressed sequence tags (ESTs)
  • Analysis of unknown sequences
  • Resequencing and comparison with reference sequences

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