DNA Sequencing Sample Submission Guidelines
For rapid processing of your automated DNA sequencing samples and to obtain the
best results, please follow these template and primer submission guidelines.
All Samples and Services
- Sequencing templates should be provided as purified plasmid, BAC, or PCR product
- Plasmid prep, PCR purification, and BAC culturing services are available, if needed
- Dissolve templates in water only; other buffers may inhibit the sequencing reaction
- Samples should 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
|
|
BAC (>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)
|
10 µl for 1st reaction + 5 μL per add’l reaction
|
|
Primer (BAC)
|
10 μM (pmol/μL)
|
10 µl for 1st reaction + 5 μL per add’l reaction
|
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 SimpleSeq™ tubes (for template) or 1.7 mL microfuge tubes (for
template or primer)
- Clearly label each tube for easy identification
- We recommend using our free bar-coded labels on each tube to ensure unique identification
DNA Sequencing Templates and Primers in Plates
- Please use 96-well, v-bottom, full-skirt PCR plates
- Seal the plate with strip caps or a cap mat to prevent cross-contamination and sample
loss
- Two wells on each plate should be left empty for our internal quality controls
- There is a 48 sample minimum per plate
- A bar-coded label can be applied to the plate to ensure unique identification
- For optimal results, normalize the template concentrations across the plate to match
the concentration ranges listed above
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
Express Plate DNA Sequencing Services
- Use capped 96-well PCR plates as specified above
- Please attach a bar-coded label to each plate
- Ship samples overnight at room temperature
For 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
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.
Below, you will find a list of those most commonly used. A complete listing can
be found on
the Standard Primer List page . 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.
Partial List of Standard Primers
|
|
Vector Primer
|
Primer Sequence
|
Vector Primer
|
Min. Volumes
|
|
M13F
|
TGT AAA ACG ACG GCC AGT
|
T7Term
|
CTA GTT ATT GCT CAG CGG T
|
|
M13R
|
CAG GAA ACA GCT ATG ACC
|
pBadF
|
ATG CCA TAG CAT TTT TAT CC
|
|
M13F43
|
AGG GTT TTC CCA GTC ACG ACG TT
|
pBadR
|
GAT TTA ATC TGT ATC AGG
|
|
M13R49
|
GAG CGG ATA ACA ATT TCA CAC AGG
|
petup
|
ATG CGT CCG GCG TAG A
|
|
SP6
|
CA TTT AGG TGA CAC TAT AG
|
pGexF
|
ATA GCA TGG CCT TTG CAG G
|
|
T3
|
AAT TAA CCC TCA CTA AAG GG
|
pGexR
|
GAG CTG CAT GTG TCA GAG G
|
|
T7
|
TAA TAC GAC TCA CTA TAG GG
|
BGHrev
|
TAG AAG GCA CAG TCG AGG
|
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 1.7 mL tubes or 96-well, full-skirt PCR plates
- Seal with strip caps or a cap mat
- Include a gel picture, if available
- Ship overnight at room temperature
Ordering DNA Sequencing Services
To order Eurofins MWG Operon DNA sequencing services, visit our
DNA Sequencing Order Entry page. Follow the instructions to order DNA sequencing
services for plasmids, PCR products, and BACs 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
dnatechsupport@eurofins.com
Sincerely,
Your Eurofins MWG Operon Support Team
Phone: 1 (800) 688-2248
Fax: 1 (251) 252-7794
Email: dnatechsupport@eurofins.com