Sample Submission Guidelines
Automated DNA Sequencing Services
For rapid processing of your sequencing samples with the best results, please follow these sample submission guidelines.
All Samples and Services
- Templates should be provided as purified plasmid or PCR product.
- Plasmid prep, PCR purification, and large clone culturing services are available, if needed
- Dissolve templates in water as EDTA, present in TE, will inhibit the sequencing reaction
- Samples can be shipped overnight at room temperature
Recommended Concentrations & Volumes |
|
Template |
Concentration |
Min. Volumes |
|
Plasmid |
150–300 ng/µl |
10 µl for 1st reaction + 5 μL per add’l reaction |
|
PCR <300 Bp |
10–20 ng/µl |
10 µl for 1st reaction + 5 μL per add’l reaction |
|
PCR 300–1000 Bp |
20–40 ng/µl |
10 µl for 1st reaction + 5 μL per add’l reaction |
|
PCR >1000 Bp |
30–60 ng/µl |
10 µl for 1st reaction + 5 μL per add’l reaction |
| Large Clones >20 kBp |
500–1000 ng/μL |
10 µl for 1st reaction + 5 μL per add’l reaction |
| Primer (plasmid or PCR) |
2 μM (pmol/μL) |
5 μL for every 2 reactions |
| Primer (large clone) |
10 μM (pmol/μL) |
5 μL for every 2 reactions |
Please review your DNA sequencing order summary sheet for exact recommendations based on your specific order.
DNA Sequencing Templates and Primers in Tubes
- Please use 1.5 mL conical-bottom tubes.
- A bar-coded label can be attached to the tube to ensure unique identification
DNA Sequencing Templates and Primers in Plates
- Please use 96-well, v-bottom, full-skirt PCR plates
- Fill the plate by column and please do not skip wells; there is a 48 well minimum per plate
- Plate positions G12 and H12 are reserved for our internal quality controls
- Seal the plate with an adhesive foil cover so that every well is sealed
- A bar-coded label can be applied to the foil seal to ensure unique identification
- For optimal results, normalize the template concentrations across the plate
Premixed Samples with Primers
- Please make template and primer solutions that match the concentration ranges listed above
- Mix 8 μL of template and 4 μL of primer for a total volume of 12 μL
Premixed Samples with Primers
- Use either the 1.5 mL tubes or the 96-well PCR plates specified above
- Only reactions using ABI BigDye Terminator Kit (v3.1) can be accepted
- The dye terminator clean-up procedure must be performed before submitting samples
- Dried or wet samples are accepted; dried samples produce the best results
- Dried samples may be shipped overnight at room temp; wet samples must be shipped on ice
- Please attach a bar-coded label to each tube or plate
Ship DNA sequencing template and primers to:
Eurofins MWG Operon
Attention: DNA Sequencing Lab
2211 Seminole Drive
Huntsville, AL 35805
Primer Selection
Eurofins MWG Operon offers a wide selection of free standard primers for DNA sequencing. A complete listing can be found here. You may
also design your own primers or have us design, synthesize, and store your primers for an additional charge.
If you choose to design your own primers, we recommend a length of 18–22 bases, 50–60% GC content, no more
than 3 consecutive repeated bases, and an annealing temperature of 50–60°C.
Sample Submission for Plasmid Preparation
- Pellet cells in standard 1.5 mL tubes or 2 mL 96-well, deep-well plates
- Ship overnight on dry ice
Sample Submission for PCR Purification
- Dispense 10 – 50 μL of PCR product in 1.5 mL tubes or 96-well, full-skirt PCR plates
- Include a gel picture, if available
- Ship overnight on ice
Sample Submission for PCR Purification
To order Eurofins MWG Operon DNA sequencing services, visit www.operon.com, click the Order Now button
on the right, and select Order – DNA Sequencing. Follow the instructions to order DNA sequencing services for plasmids, PCR products, and large clone inserts in tubes and 96-well plates. You can also order free bar-coded labels by using the barcode management center found at the top of the DNA sequencing services ordering page.
IMPORTANT: If you do not attach bar-coded labels to your submitted templates and primers, please enclose a copy
of the online order confirmation page with every shipment to ensure proper identification, tracking, and results
retrieval. If you are using Eurofins MWG Operon bar-coded labels and have entered the barcodes during the order
entry process, no further identification is necessary. The barcodes on your template and primer tubes and plates will serve as unique identification and allow fast, accurate, and secure retrieval of your results.
If you have questions about our DNA sequencing sample submission guidelines or the online order entry
system, please call us between 7:00 am to 7:00 pm Central Time for immediate assistance or send an e-mail to seqsupport@operon.com
Sincerely,
Your Eurofins MWG Operon Support Team
Phone: 1 (800) 688-2248
Fax: 1 (251) 252-7794
Email: seqsupport@operon.com