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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
-
$28.00
for up to 1,200 bases with labeled primers;
-
$36.00 for
up to 1,200 bases with dye terminators (labeling fee included);
>1.5 Kb or Primer Walking
-
$0.06/base.
Sequencing, when possible and unless otherwise
indicated, will be from both orientations.
-
If
primer walking is required,
a $28.00 fee (1 free labeling
included) will be charged for each internal walking primer
synthesized.
-
Please
note that synthesized sequencing primers can be reused in the future. To
facilitate process, please ask for your primer reference number.
Sequencing
with dye terminators
PCR
oligos can be used for sequencing along with dye-labeled terminators. A
labeling fee of $5.00 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 future sequencing 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 local shipping fee of $10.00 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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