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Frequently Asked Questions

» Sanger Sequencing Services

Q1. What services do you offer?

We offer Sanger DNA sequencing, PCR, Plasmid Purification, Primer Walking, SNP Analysis, Capillary Electrophoresis, Picking & Growing Colonies, on already sequenced and cleaned up samples (Ready to Inject Plates).

Q2. What is the turnaround time for results?

The turnaround time depends on the number of reactions.

For individual sample orders sent as purified DNA or crude PCR product the turnaround time is normally 24 hours if samples are received by 4pm. Re-sequencing reactions for any reason adds an extra 24 hours approximately (not including weekends).

For plate orders the turnaround time can take two to three business days. For plate orders containing a large number of plates the turnaround time will be a little longer, but we will provide you the results as we get them done which should start within 72 hours.

It will take one extra day for samples sent as colonies on agar plates for plasmid purification or for pelleted cultures.

Q3. What machines do you use?

We use the ABI 3730xl DNA Sequencer with 50cm arrays.

Q4. What is the average read length per sample?

All of our reads will read out to over 1000 bases; however we typically get approximately 800 bases of high quality sequence per reaction. The read length will vary depending on sample quality.

» Sample Requirements

Q1. What concentration should my primer be?

All primers should be sent at 10µM.

Q2. What volume of primer should I send?

For individual samples a minimum of 10µl should be sent. If you are using the same primer on multiple samples, we require just 3µl multiplied by the number of reactions you are doing with that primer, but still a minimum of 10µl.

For full plates we need 100µl of primer at 10µM per plate. If you are sequencing multiple plates that will use the same primer, send 100µl plus 30µl for each additional plate.

Q3. Can I premix the sample and primer?

Yes. Be sure you have a total of 20 picomoles of primer per 15µl of premix. We recommend using the following formula to create pre-mix samples:

DNA: N=0.1 x (vector size in base pairs + insert size in base pairs)

*N is the nanograms of DNA that should be added to each 15µl of premix.

Q4. What should the sample concentration be?

The sample type will determine the concentration needed.

i. PCR Product: 25-50 ng/µl

ii. Plasmid DNA: minimum of 50 ng/µl but we prefer it at 100 ng/µl. If you are sending it at 100ng/µl we need 10µl for one reaction or 15µl for two reactions. If you are sending it at less than 100ng/µl please send 15µl for one reaction or 25µl for two reactions.

Q5. I am sending cultured pellets and am unsure if they are sufficient. Do you have any suggestions?

If you can check the O.D. of the solution before pelleting, it should be around 3.0-4.0. If you did not check the O.D. prior to pelleting , feel free to send the pellets and, if upon arrival they appear too small, we will re-grow them for you at no charge and proceed to contact you to discuss changes for future plates to help ensure proper size.

*If we have to re-grow them this will add an additional day for turnaround time.

Q6. Do I have to send my own primer?

No. We have a list of free primers. We can also synthesize and/or design your primers for you. Primers are stored for a period of 2 years.

Q7. What primers do you offer for free?

Please refer to the primers page on the website for a complete list.

Q8. Can you do primer synthesis?

Yes. It is $16 per primer for standard primers. If you want us to design and synthesize the primer the cost is $25 per primer.

» Sample Submission

Q1. How do I create an account?

Please visit our account registration page and follow the prompts.

Q3. Can I modify my order after it has been submitted?

No, you cannot. If you have not yet sent your samples you can record changes to the order on the copy of the order form you printed out and are sending along with the samples. If you already sent the samples you can email us at sequence@functionalbio.com to explain the change or call us at (608)441-8125 / (877)293-7947.

Q4. When creating my sample submission form, I would like to attach a file indicating specific instructions. Would you prefer this in a specific format?

We offer a template on our website that you can use for your convenience. If you are sending individual samples use either the PCR product spreadsheet or the plasmid spreadsheet. On the sample submission form, when entering the sample names, click on the box that says "check here to import sample information in a batch" and then copy the yellow cells from the spreadsheets.

For full plates use the full plate template you can save a spreadsheet for each plate you are sending and then upload it in the box next to the plate name on the order form.

*Please be sure not to use any characters other than letters, numbers and dashes/underscores.

Q5. What if I already submitted my order and forgot to include a spreadsheet?

If you already submitted the order feel free to email us a copy of the spreadsheet, sequence@functionalbio.com, and indicate which order it should go with.

Q6. Can I cancel my order?

Yes. If you do not send the samples, we will not process the order.

Q7. Which carrier should I use for shipment?

Whichever is most convenient for you; we recommend UPS or FedEx for faster shipping. It is best to send the samples priority overnight so we get them by 10:30 a.m. If you send them standard overnight, we may get them in the morning but sometimes we do not get standard overnight shipments until 3 p.m. which can delay the results. If you choose to send samples USPS, please understand that there will be an extra day for turnaround as we do not receive those packages until late in the afternoon.

Q8. Can I request a custom annealing temperature for my samples?

Yes, but requesting custom annealing temperatures will cause a delay in sequencing results or an additional charge to the order if requested for re-sequencing.

» Sequencing Results

Q1. How do I receive the results?

We will send you an email when your results are posted to the website. You can then login and download your results.

Q2. Will you keep my samples and/or primers?

We will hold on to both for approximately two months unless otherwise requested. If we synthesize a primer for you we will put it in long-term storage.

Q3. Can I get my samples back?

Yes. We will ship them to you if requested; however, we will add the shipping charge to your invoice for this service.

Q4. Can I submit a new order and request to use samples I previously sent to you?

Yes. Please call or email to be sure we still have the samples you are requesting.

» Billing

Q1. What are my payment options?

You can pay by check, credit card, wire transfer, or direct deposit. We accept Visa and MasterCard (online only). We can also set you up with a pre-paid account or automatic payments.

Q2. When can I expect to be invoiced?

Invoices are sent out every Tuesday and Thursday. You can expect an invoice after you have received you results.

Q3. Do you require a PO on all orders?

No, we do not require a PO. This is up to your institution. If you are required to have a PO by your institution, please indicate the PO number on your sample submission form along with the address that the invoice should be mailed to.