How to Submit Samples

For best results:

Sequencing templates should be provided as purified plasmid, BAC, or PCR product
Suspend templates in water; buffers containing EDTA will inhibit the sequencing reaction.

Premixed Reactions

New! SimpleSeq™ Webless™ pre-mixed reaction submissions

Prepare aqueous template and primer solutions at the concentrations listed below.
Add 8 μL template + 4 μL primer to each tube by inserting your pipet tip through the split seal (total volume =12 μL).

Submitting standard or SimpleSeq™ pre-mixed reactions

Prepare aqueous template and primer solutions at the concentrations listed below.
Place a barcoded label on any tubes without a barcode. SimpleSeq tubes will be pre-labeled.
Add 8 μL template + 4 μL primer to each tube (total volume =12 μL).
4. Enter the details of your reactions in our online order form.

Express Plate™ DNA Sequencing

Prepare aqueous template and primer solutions at the concentrations listed below.
Using 96-well, v-bottom, full-skirt PCR plates, add 8 μL template + 4 μL primer to each well (total volume =12 μL).
Cap each plate securely and attach one bar-coded label to every plate for unique identification. Enter your order online.

Template type	PCR <300 Bp	PCR 300 – 1000 Bp	PCR >1000 Bp	Plasmids all lengths	BAC >20 KBp
Concentration	10-20 ng/μL	20-40 ng/μL	40-60 ng/μL	100-200 ng/μL	500-1000 ng/μL
Volume	10 μL for first template + 5 μL per template for subsequent reactions

Primers	PCR <300 Bp	PCR 300 – 1000 Bp	PCR >1000 Bp	Plasmids all lengths	BAC >20 KBp
Concentration	2 μM	2 μM	2 μM	2 μM	10 μM
Volume	10 μL for first primer + 5 μL per primer for subsequent reactions

Submitting Templates and Primers

Submitting templates and primers

Prepare aqueous template and primer solutions at the concentrations listed above
Place a barcoded label on any tubes without a barcode. SimpleSeq tubes will be pre-labeled.
For each template & specific primer, pipet the appropriate volume. Free standard primers can be added by our lab.
Enter the details of your reactions in our online order form.

Plate submissions for templates and primers

Samples should be shipped using 96-well, v-bottom, full-skirt PCR plates (min. 48 reactions per plate).
Please attach one barcode label to each plate for unique identification.
Two wells on each plate should be left empty for our internal quality controls at positions H11/H12 for row layouts and G12/H12 for column layouts. Enter the detail for your reactions in our online order form.
Only strip caps or cap mats should be used for sealing (no foil or tape-seals, as sample loss frequently occurs).

How to Ship Samples

Overnight services offered at the same price as standard service - No additional fees!
Order your Overnight Kit.
Each kit contains pre-addressed, personalized UPS labels and envelopes, barcode labels, and sample bags.
Please ship templates and primers at room temperature via UPS using our Overnight Kit.
If you’ve submitted an online order, please enclose a copy of the confirmation page with your shipment.

Primer Selection

Primers synthesized or shipped to us for sequencing should be between 18 and 22 bp in length, avoid stretches of identical nucleotides, display about 50% GC content, and exhibit an annealing temperature of 50–60°C.
Primers that are synthesized for your sequencing order remain available to use for six months.
Primer walking, primer design, and primer synthesis services are also available.

Free standard primers

Eurofins MWG Operon offers an extensive selection of standard sequencing primers for many common vectors including M13F, M13F43, M13R M1R49, SP6, T3, T7, T7Term, pBadF, pBadR, petup, and many others. Click here to view the full list.

Express Plate Ready-to-Load Service

Only reactions using ABI BigDye Terminator Kit (v3.1 or v1.1) can be accepted.
The dye terminator clean-up procedure must be performed before submitting samples.
Dried samples are preferred.

Plasmid Preparation Service

Glycerol Stocks Recommended: send a minimum of 10 μL of a 50% glycerol solution prepared from overnight cultures. Glycerols stocks, pelleted cells, overnight cultures, or plates should be shipped with ice packs for overnight delivery.

PCR-derived Templates

Sample submission for PCR purification or standard dilution without purification

Dispense PCR product as detailed below in 1.5 – 2.0 mL microfuge tubes or 96-well, full-skirt PCR plates.
Only strip caps or cap mats should be used for sealing (no foil or tape-seals, as sample loss frequently occurs).

	Submit	Notes
Crude PCR	entire reaction volume	15 µL minimum volume
PCR products	10 µL for standard dilution	request Power Read for dilution protocols

Online ordering for Overnight DNA sequencing

Click the blue “Order Now” button on the right of your screen
Select “DNA Sequencing” and follow instructions for Tube Entry, Tube Upload, Plate Entry, or Plate Upload ordering.

Click for free Overnight Kit or barcoded labels.

Questions? Please call us at 1(800)688-2248 between 7:00 am to 7:00 pm Central Time for immediate assistance or send an e-mail to dnatechsupport@eurofins.com

Click here for a printable copy of these guidelines

Eurofins MWG Operon > Services > DNA Sequencing > How-to-Submit Samples