How to send your samples

TEMPLATES                                                                               

For each reaction, we need 5 µl of template containing:

> 250-500 ng of plasmid DNA, i.e. at a concentration of 100 ng/µl,
> "a visible band" of purified PCR product (approx. 2 ng per 100 bp),
> 1 µg for large plasmids (cosmids or BACs) or
> 1-2 µg of bacterial genomic DNA.

Minimal volume is 5 µl: lower volumes may evaporate. Minimal plasmid concentration is 50 ng/µl.
If you have enough DNA, please provide more than 5 µl per reaction, e.g. in case we need to repeat a reaction due to difficult DNA structures.
We prefer if you provide one tube for each template (and not one tube for each reaction), with primers in separate tubes. However, large number reactions should be provided in strips or plates. For plates, the orientation of samples must be A1, B1,.., H1, A2, ..

As the DNA template is one of the main factor influencing sequence quality, it should be purified using commercial kits. 
It should be provided in distilled water (or Tris), free of EDTA and other salts,  proteins, RNA, or genomic DNA. 
PCR products have to be purified from reaction buffer, primers and nucleotides. They must appear as single band in an agarose gel.

If necessary, we can purify your DNA templates. Please contact us, as special condition applies.

PRIMERS

We supply  standard primers free of charge.

If you are not using our standard primer, please include your own primer in a separate tube at 10 µM in distilled water. We need approximately 1 µl of primer per reaction.

Ordering

Please do not hesitate to contact us for testing our service.
First time users will need to register to get access to the DNA Sequencing Order Form.
The links are found on the home page.

Data delivery

Sequencing data is delivered by e-mail.
The sequences are saved under FASTA format in text files.
The chromatograms are saved under ABI format compressed in a WinZIP archive.

Example of a control reaction, pGEM vector sequenced with the universal primer M13 Forward:

• Sequence, FASTA format
• Chromatogram, AB1 format for Windows

NB: the quality of the sequence depends significantly on the template and the primer (e.g. base composition, purification).

 The chromatograms can be visualized using the following free programs:

• FinchTV (Mac OS X, Windows, Linux, Solaris), can show quality values
• 4Peaks (Mac OS X)), can show quality values
• Sequence Scanner Software (Windows), ABI's reader, can show quality values
• BioEdit (Windows)
• ApE- A Plasmid Editor (Mac OS X, Windows, Linux)
• Sequencher Demo (Mac OS 9 or X, Windows)
• Chromas LITE (Windows)