Sanger sequencing services

FAQs

What types of Sanger sequencing services do LGC, Biosearch Technologies offer?

We offer both single sample and MTP (96-well plate) sequencing which are run as automated and standardised ABI 3730xl sequencing runs, and obtain read lengths up to 1,100 nucleotides (nt) (PHRED20 quality).

The following standard service options are available:

• Ready2 Run - service: sample and primer pre-mixed by customer
• Flexi Run - service: sample and primer are sent separately by the customer and will be stored for 3 months with the possibility to re-use them. We offer a set of standard primers, custom primers can submitted, or synthesis of specific primers can be ordered.
• Premium - service: Here we perform a sample DNA quality check first to find optimal sequencing conditions. We can adapt sequencing conditions further for samples such as large constructs (e.g. BACs), low concentration samples, or GC-rich samples. PCR clean up and plasmid preparations are included for MTP sequencing and can be ordered optionally for single sample sequencing.

Individual projects are also possible to reflect your specific requirements. We can offer additional services including complete 16S/ITS analysis, primer walking, BOL, re-sequencing. Please feel free to discuss your project with us.

What sample types can be submitted for Sanger sequencing services?

PCR products and plasmids (up to 12 kB) in our standard service options (Ready2/Flexi-run) and cosmid only in Premium Services.
Please note that we cannot accept genomic DNA.

What is the required concentration for submitted DNA?

The concentrations must be adjusted prior to shipping for the standard service options (Ready2/Flexi-run)

PCR products:

• 150-500 bp: 10 ng/µL
• 500-1000 bp: 20 ng/µL
• 1000-2000 bp: 40ng/µL

Plasmids:

• 100 ng/µL

Please note that for MTP services, PCR products should not differ in size and concentration by a factor of 3 within the plate as homogeneous sequencing conditions must be used across all samples in the plate.

Is there a DNA quality requirement?

For our standard service (Ready2/Flexi-run) submitted DNA samples must be purified by the customer. For our Premium Service, sample purification (enzymatic) can be selected as an option when ordering.

We recommend eluting the samples in water to avoid the presence of inhibiting residues in the sample. Sanger sequencing primers and templates need be free from fluorophores (no fluorescent FAM-/HEX-/... modification) as their presence disturbs the analyses and can potentially cause severe damage to the sequencing instruments.

What recommendations do you have regarding primers?

There are three options for primers in Sanger sequencing services:

1. Use standard primers – these are universal primers that we offer in-house. A list of primers is available in your online account.
2. Customer-submitted primers – submit your preferred primers with your samples (combined together for Ready2 Run services, and separately for Flexi Run services).
3. Order synthesis of required primers - we can synthesize primers for you (design of primers is available only with our premium service). Primer synthesis should be ordered before 3 pm (working days only) to guarantee sequencing on the next working day.

Customer-submitted primers and in-house primer syntheses are stored for 3 months.
We recommend providing primers as stock solutions (100 μM).
For your calculations: 4 μL of a 5 μM primer are required for each sequencing reaction. Please ensure that excess primers are supplied to enable any repeat reactions to be perfomed.

What is the estimated turnaround time for regular sequencing requests?

Once the samples have arrived, they will be processed immediately. Results are available within approximately 24 or 48 hours, depending on the selected service.
 
We also offer an express service (for Germany and Austria). If this has not yet been established at your location, please contact our sequencing support team (sequencing@lgcgroup.com) for further details.
 
For individual customized projects, the processing times are coordinated and communicated with the responsible project manager.

What sample shipment option should I use?

There are several alternative options for sample shipment including courier, standard mail, or a pick-up box.

Please check with our sequencing support team (sequencing@lgcgroup.com) to determine which option would be the most suitable for your location.

Courier forms for MTP (96-well plate) shipments are available in your online account.

How do I get started?

Please create a customer account in our Sanger Sequencing Online Shop. Once the account is activated, please order a batch of barcode labels that are essential for our sequencing service (these are free of charge). Additionally, you can order free sample bags and/or MTP shipping boxes for safe shipment of your templates.

For submission of sequencing requests please contact our sequencing support team (sequencing@lgcgroup.com).

What are the payment options for Sanger sequencing services?

We offer a credit system for invoicing of sequencing orders, and this can be used for a range of our sequencing services.

You have the option to load a specified amount (minimum 50 Euros) into your Sanger account. This credit is available to use immediately, and you can start ordering Sanger sequencing. You will subsequently receive an invoice for the amount loaded into your account.

Furthermore, you can mention a PO, or respectively, a project number in online ordering for improve traceability for your financial department.

Alternatively, sequencing orders can be paid individually per invoice. It should be noted here that the prices per sequencing are higher when individual invoicing is used, compared to the pre-loaded credit option.

How will I be updated regarding the status of my sequencing request?

You will regularly be informed about the status of your request once the templates have arrived at our facility. These communications will be in the form of automated emails to inform you of sample receipt, data availability etc. Furthermore, the current state of progress is mentioned in the section ‘Data management’: ‘Previous orders & download results’ section of your online account.

How do I go about interpreting my Sanger sequencing results?

The data sets are available for download in your online LGC sequencing account for 3 months (section ‘Data management’: ‘Previous orders & download results’).

We strongly recommend checking the trace files (electropherograms) from the sequencing when performing any analysis, rather than relying solely on the sequence itself. Trace data from Sanger DNA Sequencing (.scf or .ab1 file formats) can be displayed by freeware programs including FinchTV or Chromas lite. All achieved datasets are Phred20 quality clipped.

How do I seek help with troubleshooting?

If you have any questions about your data, please contact our sequencing support team:

Tel: +49 (0)30 5304 -2230 | - 2298 | - 2233

Email: sequencing@lgcgroup.com