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DNA Sequencing
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Welcome to our Bioanalysis Division. You
need fast and reliable DNA sequencing, this is the service you need. Should
you have any questions or requests, please don't hesitate and contact
us.
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Fig 1. Fluorescence-based dye-terminator sequencing chemistry. Fluorescent
signal in a "chromatogram-like" format.
Service Offer
Single pass sequencing service
Basic Service, for quick and economical sequence determination.
Features:
• 400-600 bases read depending upon sample quality
• Plasmids, PCR fragments, Lambda
• Universal primers included (M13, T3, T7, SP6, pBluescript, pGEM,
etc.)
• Results through E-mail and hard copy by mail
Single-strand sequencing service
Most economical approach to sequencing long fragments ( < 7kb)
when the data is not intended for publication. Features:
• Sequencing is performed on one strand only
• Plasmids, PCR fragments, Lambda, Cosmids
• Primer walking approach
• Contig alignment, edition.
• All primers included
• E-mailed results + Complete written report
Double strand sequencing publication quality service
• Double-strand sequencing
• Plasmids, PCR fragments, Lambda, Cosmids, BACs
• Walks / shot gun
• All Ambiguities resolved
•Guaranteed results
•Contig alignment, edition and consensus sequence determination
•All primers included
•E-mailed results + Complete written report (RE, ORF, Protein, codons...)
Sample Requirements
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. Your DNA should be:
(i) accurately quantified
& (ii) adequately prepared DNA for enzymatic sequencing reactions
See troubleshooting
Purification and 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 H2O at a minimum concentration of 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.
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 H2O and quantified by UV at 260 nm.
TE buffer should be avoided.
Lambda, Cosmid, 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 H2O at a minimal concentration of 0.5 µg/µl.
Amounts Required:
| Insert size |
Approximate
Amount
For Single-Pass Sequencing |
Approximate
Amount
For Full Length Sequencing |
| Plasmid Up to 5 kb |
2 to 3 µg / reaction |
5 µg / 1.5 kb |
| PCR product Up to 3 kb |
0.5 to 1 µg / reaction |
1 µg / kb |
| Cloned 5 to 10 kb |
5 µg |
30 µg |
| Cloned 10 to 20 kb |
5 to 10 µg |
50 µg |
| > 20 kb and BACs |
Please contact us |
Please contact us |
Provided Primers
Only applicable to Single-Pass Service. Please, no degenerate primers.
Provide us with: 10 to 20 µl of a 10 µM (10 pmol/µl)
aqueous solution of your specific primer(s). Please indicate sequence
of primer(s) sent.
Final note
Samples are not returned and will be kept one month following your order
before disposal.
Ordering
and Shipping of your Samples (see also General Ordering Information)
Please send your tubes, clearly labeled and wrapped with
parafilm in a padded envelope. Include your Institutional Purchase Order
and a letter describing your samples and indicating desired primers for
Single-Pass sequencing, flanking sequences or primers in all other cases:
1- Sample name, Vector, Insert size (if known), Total amount (µg),
Concentration (µg/µl) Volume (µl) and solution (H2O,
buffers...).
2- Sequencing primers
Send using regular Postal Services at:
GENOSPHERE BIOTECHNOLOGIES
DNA Sequencing
2 rue des Gravilliers
75003 PARIS
FRANCE
Receipt of your samples is confirmed to you by e-mail and an approximate
date of completion is released to you.
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