Sanger Sequencing FAQs
What payment methods can I use when ordering a Sanger Sequencing job?
Source BioScience can process the payment of sequencing orders through purchase order number, eVouchers or credit card.
What eVouchers are available to order and how can I order them?
Source BioScience offers a range of single read and 96 well plate eVouchers. Click on the following link to find out more and order your Sanger Sequencing eVouchers.
How many eVouchers are required per job?
Each reaction that is ordered on the website requires one eVoucher to cover the cost of the reaction. When you get to the ‘your basket’ page on the order form simply input the number of eVouchers you are using on the job and click update cart in the bottom right-hand corner. This will subtract the cost of the eVouchers from the order. eVouchers are automatically deducted from your account. If you are submitting a full plate for sequencing then 96 single read eVouchers or 1x 96 well plate eVouchers will be required as payment.
NOTE: eVouchers cannot be used to pay for PCR clean-ups, template amplification, primers, dGTP, Colony PCR or Bugs2Bases. These treatments require a purchase order number or credit card as payment.
Can I use eVouchers to cover the cost of a Colony PCR order?
You can use eVouchers to cover the cost of the Sequencing reaction, but the Colony PCR reaction will need to be paid for using a PO number, Credit Card or Spenddown account.
How much DNA and Primer do I need to send in for a Sequencing Order?
The sample requirements for Sequencing can be found in the following link DNA and Primer Sample Requirements.
How do I order a primer with my Sequencing Job?
Primers can be ordered on the 4th page of the Sequencing Online Order Form. Simply add a primer name and the primer sequence into the designated fields and click ‘add this primer’. Continue with the order as normal and input a purchase order number in the ‘checkout’ page at the end of the order. Once the order has been confirmed your primer synthesis request is immediately placed and will arrive with us quoting your sequencing job number.
How long do you store DNA and primers for?
Customer DNA and primers that are submitted to us are stored for 3 weeks before being discarded.
How long do you store synthesised primers for?
Primers that are ordered through the website are stored for a year and can be selected from the drop-down box on the reaction details page.
How do I request the re-use of DNA and primers that have already been submitted?
The first question on the Online Order Form asks if you want to re-use DNA or primers from a previously submitted order. Select ‘Yes’ and then fill out the rest of the fields below, remembering to input the previous order number.
Do my Colony PCR primers need to be sent in separate tubes?
Yes, all primers we process need to be sent to us in separate tubes so that we can prepare the Master Mixes with the correct ratio of reagents and primer.
Can I use the same primers for Colony PCR and Sequencing?
Yes, although we require primers sent at different concentrations for the Colony PCR and Sequencing. Our primer requirements can be found here
How do I access the List of Primers that Source BioScience Stock?
You can check what primers Source BioScience stock by clicking here.
I have been unable to generate the amount of DNA required for Source BioScience. Can I still send my samples?
If you are unable to generate the volume and concentration of DNA we require then please contact the laboratory or our sales team who can advise you whether it is worth sending the samples. The sequencing reactions can be optimised to work with stronger/weaker DNA.
What are the different reaction types?
The pre-sequenced reaction service requires customers to supply a purified sequencing reaction that is ready for processing on the sequencer. The DNA and primer will have been PCR cycled and purified before sending to SBS for data collection.
Sequencing Reaction Clean-up
The sequencing reaction clean-up is the reaction purification prior to samples being processed on the sequencer. The DNA and primer will have been PCR cycled but NOT purified before sending to SBS for data collection.
PCR Reaction Clean-up
The PCR reaction clean-up service is the purification of PCR products sent to SBS for sequencing.
The template amplification service is only suitable for plasmid DNA. The service involves a 12-hour incubation period to increase DNA concentration and improve DNA quality by removing any inhibitors that could impact the sequencing reaction.
Analysis Settings and Data Options
What is the difference between the SpeedREAD™ automated data delivery service and the attached email service?
The SpeedREAD™ service collects the data as soon as it has finished on the sequencer and processes the data through a QC procedure, looking at the length and quality of the traces. If the data passes this QC step, an automatic email containing a hyperlink to the data is sent to the customer. When SpeedREAD™ is selected we aim to have the data sent back to each customer by 9am the next morning. This method also helps to control the size of the email that is received by allowing the data to be downloaded by a hyperlink contained in the email.
For an in-depth description of the SpeedREAD™, please click here.
The attached email service is a manual process which includes a member of the sequencing team analysing the data and dispatching to the customer via an attachment to an email. The emails tend to be larger due to the attachments.
What is the Turnaround Time for Colony PCR orders?
We aim to return all Colony PCR data within 24 hours of sample receipt.
How long do you keep Colony PCR samples and primers?
We keep the Colony PCR samples and primers for the standard 3 weeks before discarding them. If you would like us to store them for longer then please contact us to request this ([email protected])
What is the Basecaller used by Source BioScience?
For an in-depth description of the PeakTrace™ Basecaller, please click here.
What is Secondary Structure Resolution?
Source BioScience provides a chemistry known as the dGTP chemistry to help resolve sequencing issues such as G-C rich templates, hairpin structures, repetitive regions or troublesome sequences. The dGTP chemistry can be selected on the online order form. There is an additional £3 charge per reaction for this chemistry and it can only be paid for by credit card or purchase order number.
What is Quality Value Limit, Mixed Base Identification and Quality clipping?
For an in-depth description of the quality value limit, mixed base identification and quality clipping selections then please click here.
What format do the data files arrive in and which programs can I use to open them?
Each sequence reaction produces 2 files in different formats, the .ab1 file and the .seq file.
The .ab1 file contains the electropherogram and raw data traces and can be opened in most sequencing software, click here to be directed to a website for downloading the free software FinchTV.
The .seq file contains the base calls from the sequencing reactions which can be opened in notepad, word pad or word document.
What should my Sequencing Data look like?
For a detailed description of the different types of sequencing traces generated please click here.
I am using a general lab account but would like the data to be emailed to me too; how do I add an additional email address to the order?
Additional email addresses can be added at the bottom of the ‘Analysis Settings’ page on the online order form. If you are inputting more than one email address please separate with a semi-colon (;).
General Sequencing Order Queries
I have selected on the order form to send the samples by post but I am actually using a collection point, do I need to change the order?
No action is required, our collection points are visited whether an order has been selected for pick-up from that site or not, so we will still receive the order as normal.
I made a mistake on the online order form? Do I need to place a new order?
If an order has been placed that contains incorrect information then please contact us here stating the order number affected. We will reject the order from the system and ask you to place another order in its place.
The data I received isn’t as good as I expected. Can I get the reactions repeated?
We offer a repeat sequencing service for failed runs from samples which meet our submission requirements. If you receive reads that fail to generate sequences as per your expectations, please reply to the email within which you received the data and list the reaction(s) that you would like repeated. Any background information on size of the fragment and sample type can help us troubleshoot the data. Our lab team will analyse the submitted samples to verify that the sample requirements have been met. If the samples are found to meet our requirements, the sequencing will be repeated, free of charge.
Should your samples fail to meet the requirements, Source BioScience will offer a courtesy repeat sequencing service on up to 10% of the samples, to see if the deficiencies can be adjusted for. You will be made aware of the failed reads immediately and will be given the option to determine which samples, if any, will undergo the repeated sequencing.
You can contact us here to request the repeats you require.
I wish to discuss my data with the sequencing team, how do I contact them?
You can contact the lab that processed your reactions by either replying to the data email that contained your data files or contacting the lab using the contact details at the bottom of the data email.
Your website will not accept my order, what do I do?
The website has a limit of 12 characters for sample names and primer names, so if the names you are entering are longer, the website will not allow you to proceed with the order.
You can only input certain characters on the website, so please use only underscores (_), letters and numbers.
Full stops (.), dashes (-), brackets (), slashes (\/), stars (*), question marks (?), hash symbols (#) and commas (,) are not permitted on the order form.
For further information and prices please contact us or call +44 (0)115 973 9012