• Synthesis of single-stranded RT-PCR from an RNA template via reverse transcription using a commercially available kit.
  
  
• Synthesis of double-stranded RT-PCR from an RNA template via reverse transcription using a commercially available kit.

PCR Primer Design and Synthesis

The sequence data provided is used to design the appropriate primers required for the PCR reactions and subsequent sequencing. All primers are optimised, and their physical properties—such as GC content, Tm and self-annealing—are verified by our team of experts. Primers are then synthesised in-house.

PCR and Purification

The PCR reactions are performed on standard genomic DNA (where available) or, alternatively, on one or two of the genomic DNA samples provided by the customer. The PCR products are then purified and subjected to quality control via agarose gel electrophoresis. Any PCR products with unsatisfactory results are regenerated in a second and, if necessary, a third round. This involves a new primer design for each PCR product, primer synthesis, PCR amplification and purification of the PCR products.

Sequencing of the test PCR

To ensure the quality of the PCR process, all established PCR reactions are subsequently sequenced from both strands (bidirectional).

High-throughput PCR and purification

Once the PCR primers have been successfully designed, PCR reactions are prepared using all DNA templates and carried out using a high-throughput method. The PCR products are then purified.

High-throughput sequencing

All PCR products are sequenced in double-stranded or, on request, single-stranded quality. This step includes repeating the sequencing reaction should a reaction fail. If both reactions fail, the sample is repeated from the start.

Bioinformatics involves the high-quality trimming and assembly of individual reads, as well as the analysis of sequence data for individual gene segments. Upon request, we use the resulting consensus sequences to perform a database search (BLAST) to determine the biological classification of each sample.

We compare each sequence with a reference sequence provided by the customer and analyse it to identify potential sequence differences such as heterozygotes, insertions/deletions or codon changes.

To generate a comprehensive mutation report, we carry out an analysis using appropriate software.

All results are summarised in the form of a Certificate of Analysis (CoA) / a comprehensive report and provided alongside the raw data.

You will receive the following raw data in your online account:

• *.ab1: Electropherograms in AB1 format
• *.cons: Consensus sequence in FASTA format (MS-DOS TXT)
• *.qual: Quality report for each consensus sequence (MS-DOS TXT)
• *.align: Alignment of individual reads to generate a consensus (MS-DOS TXT)
• *.comp: Alignment comparison with a reference sequence (MS-DOS TXT)
• *.mutation: tabular summary of all observed differences from the reference sequence (XLS or MS-DOS TXT)

Certificate of Analysis (CoA)

A formal document produced by a laboratory summarising the results, specifications and methods of one or more laboratory analyses. (Bioinformatic analysis required)

Comprehensive report

A complete, detailed report in accordance with your specifications, referencing ISO 17025 accreditation, containing information on materials, methods and results. (Bioinformatic analysis required)

Sequencing is performed according to standard methods and in compliance with:

CLSI Guideline MM09-A; Vol. 34 No. 4, 2014; Nucleic Acid Sequencing Methods in Diagnostic Laboratory Medicine, Approved Guideline-Second Edition

Conduct of sequencing studies is performed according to

OECD Guideline ENV/MC/CHEM (98)17 (rev.1999): The Principles of Good Laboratory Praxis (Monograph No. 01) and OECD Guideline ENV/JM/MONO(1999)23 (rev. 2002): Short Term Study and GLP (Monograph No. 07)

Quality assurance (QA) activities related to sequencing studies (review of the study plan, inspection of study, study report and raw data) are conducted according to

OECD Guideline ENV/CBC/MONO(2022)20: Quality Assurance and GLP (Monograph No. 23).

Typical applications for Re-Sequencing / Sanger Sequencing Projects are:

 ✔️ Re‑sequencing of viral, chromosomal or complex DNA regions

 ✔️ RNA‑based sequencing projects (mRNA, viral RNA, total RNA)

 ✔️ Confirmation of sequence integrity after process changes

 ✔️ Investigation of unexpected variants or mutations

 ✔️ Support of regulatory submissions and audits

 ✔️ Long or problematic regions not covered by routine workflows

 ✔️ Dedicated project support from start to finish

Personalized guidance by an experienced project team at every stage of your sequencing project.

 ✔️ Broad sample acceptance

Processing of sample material up to biosafety levels BSL1/S1 and BSL2/S2.

 ✔️ Primer design and synthesis included

End‑to‑end support — from sequencing strategy to primer design and synthesis.

 ✔️ Advanced Bio‑IT analysis options

Comprehensive bioinformatic solutions including sequence clipping, assembly, consensus generation, and mutation analysis:

• Compilation of sequencing data into validated consensus sequence
• Quality control of the consensus sequence including detailed quality reports
  
  

 ✔️ Flexible data delivery

Multiple data formats and delivery options tailored to your workflow.

 ✔️ Secure short‑term storage included

Free storage of template DNA and all primers for up to 3 months.

 ✔️ Exceptional sequencing accuracy

Verification accuracy of up to >99.99999%, powered by the Sanger gold standard.