DNA sequencing-USA

Canadian customers please click here

To download chromatogram-viewing software: CLICK HERE


Turnaround time

Turnaround time for single pass is 24 hours. Typically, and when sequencing from both orientations, turnaround time is approximately 1 day for <1.8 Kb; approximately 4.0 Kb per week for insert sizes larger than 1.8 Kb.

Deliverables

Client will receive sequencing data (in text format) via e-mail. Chromatograms may be provided by e-mail or mail. Please print contact information clearly.

Data quality

For a sample of acceptable quality and quantity, an accuracy of >99.00% (Li-Cor standards and edited) is guaranteed for single-strand sequencing. 100% base pair matching is guaranteed for double-strand sequencing. All single-strand sequences are edited to ensure accuracy.

Fees and conditions

Single pass

· $25.00 for 1 reaction;

· $31.00 for <700 bases with dye terminators (labeling fee included);


>1.5 Kb or Primer Walking

· $0.10/base. Sequencing, when possible and unless otherwise indicated, will be from both orientations.

· Please note that synthesized sequencing primers can be reused in the future. To facilitate process, please ask for your primer reference number.

· No charge for internal walking primer (only for inserts larger than 1.5Kb)!


Sequencing with dye terminators

PCR oligos can be used for sequencing along with dye-labeled terminators. A labeling fee of $6.00 will be applied per reaction (whether primers are client-provided or synthesized by Bio S&T). We also carry many commonly used vector primers (please contact us for details). Should a vector primer not be available, we will synthesize it at no cost.

Sequencing with labeled primers

Samples will be sequenced with our labeled universal primers (T7, T7t, T3, M13F, M13R, BGH, KS, SP6, etc.). Clients who have provided labeled custom primers and wish to re-use them in futuresequencing reactions, will not be charged a labeling fee.

Editing fee

No charge for single-strand editing. For double-strand editing, the two complementary strands will be edited against each other pair by pair. An editing fee of $7.50 per 500 bases per strand applies.

Difficult template

When the template contains highly repetitive sequences (dinucleotide repeats, polynucleotides), high G-C or A-T content, or very strong secondary structures that will affect the normal sequencing procedure, additional processing fees might be added (following client’s consent) to the regular price to a maximum of 25% of the total cost.

PCR amplification

Sometimes amplification of a region prior to sequencing is necessary in order to enhance a faint signal. Each PCR costs $15.00 and is performed occasionally and mainly for large inserts, double-stranding sequencing, difficult templates, etc. This step is performed (following client’s consent) when it’s considered to be more cost-effective than primer walking.

Pick-up service

Please contact us at 514.633.6006 for our FedEx account number 24 hours prior to pick-up. A shipping fee will be charged. This fee is waived for an order of $250.00 or more before taxes.

Sample Requirements and Recommendations

Template

Preparation: Use of Qiagen kits, phenol-chloroform extraction or CsCl-density gradient centrifugation is recommended. If total size (insert + vector) is >7 Kb, a maxi-prep is recommended. Purity: A 260/280 ratio and an agarose gel picture of digested and undigested DNA samples should be provided. Shipping conditions: Plasmid DNA can be sent in dry form, in water or in a buffer solution. PCR products must be submitted in a dry state or in 70% ethanol to prevent degradation.

Sample Quantity and Concentration

Requested quantity of template for a single reaction is approximately 0.3 picomoles. The corresponding amount is about 200ng/1Kb of total size (insert + vector). Alternatively, the amount in ml can be calculated using the below formula. Please note that although our read-length for a single reaction can reach up to 1,200 bases under ideal conditions, the amount of template should be calculated assuming a read-length of 700-900 bases. Also, please note that 2 to 3 times of the required amount might be required in case of repeated sequencing reactions.

0.195 x Total size (Kb) / Sample concentration [ug/ul] = ul of sample needed for 1 reaction

NOTE: It is recommended that the DNA concentration be estimated by running a gel (more accurate than a spectrometer reading). Ideal concentration is >0.5 mg/ml. If concentration is too low this will result in a blank signal or the signal will be lost in the background.

Client-provided Primer

Quantity of primer needed per reaction is approximately 5 picomoles. Primer design should ideally be about 50 bases upstream of region of interest. Size of primer should ideally be 18-25 bases. Clients bear responsibility for the descriptions of their sample(s).

Please contact us for our account number with FedEx prior to pick-up.

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