Fig 1. Fluorescent dye based sequencing signal in a chromatogram format.

Click on figure to enlarge

Single pass sequencing service
Basic quality service, for quick and economical sequence determination:
 600-800 bases read depending upon sample quality
 Plasmids, PCR products purified or crude PCR mixes
 Universal primers included (M13, T3, T7, SP6, pBluescript, pGEM, etc.)
 Results by E-mail

Additional services:
 Primer synthesis for sequencing your sample
 PCR mix purification

Single-strand sequencing service
Most economical approach to sequencing long fragments (< 7kb) when the data is not intended for publication.
 Sequencing is performed on one strand only
 Plasmids, PCR fragments, Cosmids, BACs
 Primer walking approach
 Contig alignment, edition
 All primers included
 E-mailed results + Complete written report

Double-strand sequencing
 Double-strand sequencing
 Plasmids, PCR fragments, Cosmids, BACs
 Walks / shot gun
 All Ambiguities resolved
 Guaranteed results
 Publication quality
 Contig alignment, edition and consensus sequence determination
 All primers included
 E-mailed results + Complete report

Sample quality
The quality of the data collected in single-pass sequencing service is highly dependent upon the quality of your DNA preparation which we do not control. Please take all necessary actions to provide us with a DNA template that is:
(i) accurately quantified
(ii) adequately prepared for enzymatic sequencing reactions

Sample quantity
Single Pass Service : sample quantities for one reaction

Sample vialing and labeling
If at all possible, please aliquot your DNA and primers into 1.5 ml microfuge tubes. We are sorry but we do not accept tube strips. DNA templates and primers should be sent in separate tubes.
Microtiter plates: 96-well plates are accepted. Please standardize DNA concentration across the wells as indicated above and cap securely the plates.
DNA templates and primers should be sent in separate plates. In addition, you should use separate plates for crude PCR mixes that require purification and ready to sequence, purified PCR fragments.

To avoid any errors on our part, we recommend that you use strictly identical sample references on tube labels and sample submission form.

Purification and DNA form
Plasmid DNA
Any commercially available column-based preparation kit will generate good template for sequencing. Following mini-prep, plasmid DNA may be precipitated or solubilized in sterile dH2O and quantified by UV at 260 nm. Minimum concentration should be 0.1 µg/µl. In case of precipitation, a 70% ethanol wash of the pellet is critical for salt removal. TE buffer should be avoided.

Purified PCR fragment
The PCR reaction mixture should be separated by agarose gel electrophoresis. The band of interest should be cut out and the DNA fragment extracted (electroelution, commercial kits, etc.).
Dissolved in dH2O and quantified by UV at 260 nm, or by running an aliquot on minigel and comparing intensity against a control ladder. Minimum concentration should be 25 ng/µl. TE buffer should be avoided.

Crude PCR mix with a unique product/band
We can purify your PCR mixture for a nominal fee prior to sequencing as long as it has a unique product. If your PCR yields multiple products, you should separate and isolate the band of interest by agarose gel electrophoresis.

Cosmids, BACs
Please use commercially available column-based kits for isolation and purification of these large DNA molecules. Protocols should be very carefully applied to avoid chromosomal DNA carry over.
Solubilized in dH2O at a minimal concentration of 0.5 µg/µl.

Amounts required and insert size

Provided primers
Provide us with: 10 µl of a 10 µM (10 pmol/µl) aqueous solution of your specific primer(s). Please indicate sequence of primer(s) sent. Please, no degenerate primers.

Final note
Samples are not returned and will be kept one month following your order before disposal.

Pricing

We offer discount pricing for larger volume order. Please contact us or use our QUOTE form for a competitive offer.

Sending your sample

An online ORDER form is available for your convenience.

Please send your tubes or plates, clearly labeled, sealed securely, and in a padded envelope. Include your Institutional Purchase Order (mandatory) and samples and indicating desired primers for single pass service, flanking sequences or primers in all other cases:
1- Sample name, vector, insert size, total amount (µg), concentration (µg/µl) volume (µl) and solution (salts, buffers…).
2- Sequencing primers (sequence, concentration, volume, and solution).
A sample submission form is available for your convenience: ORDER.

Please send at room temperature using express courier or postal services to:
GENOSPHERE BIOTECHNOLOGIES
DNA Sequencing
2 rue des Gravilliers
75003 Paris
FRANCE
Tel: 01 42 71 70 21 (local dialing)
For a faster processing of your order and safer tracking of your samples, please advise our Customer Services of your sending a sample with courier airway bill number if available: info@genosphere-biotech.com.