When Are Eurofins Sequences Usually Uploaded
- Sample Prep
- Sample Types
- Format Types
- Ship / Submit
- Results
Sample Grooming Guidelines
The preparation of various samples for ordering a sequencing service depends on the type of sample beingness submitted, the blazon of sequencing chemistry beingness ordered and the additional services being requested. Prepare your samples based on the guidelines below.
Concentration of Stock Solutions
Prepare stock solutions of purified/prepared templates and primers in ddH2O based on the size of the template being sequenced. Utilize these stock solutions to assemble samples to be submitted in tubes or plates. These guidelines do not employ to samples for Ready2Load Sequencing service.
Table 1: Concentrations of the stock solutions of primers and prepared templates
| Template Type | Size (kilo basepairs) | [Template] (ng/μL) | [Primer] (pmol/μL) |
| PCR Products, Purified | 0.100–0.300 | ten–twenty | 2–ten |
| PCR Products, Purified | 0.301–1.000 | 20–40 | ii–10 |
| PCR Products, Purified | >one.000 | 40–threescore | 2–10 |
| Plasmid, Prepared | < 1 .000 | 30 - sixty | 2–10 |
| Plasmid, Prepared | 1.0 << 6 .0 | sixty-150 | 2–10 |
| Plasmid, Prepared | 6.0 << 20.0 | 150-250 | 2–10 |
| Large Contructs (BAC), Prepared | >20 | 250–500 | 10–15 |
Note that the concentration of the Template is in ng/μL while the concentration of Primers is in pmol/μL.
Volumes of Stock Solutions for Assembling Samples
The volumes of the Sample stock solution(due south) and the Primer solution(s) depend on the type of the sequencing services required. Ensure that the stock solutions are prepared according to the guidelines in table ane before proceeding to section below.
'Premixed' Samples:
When you lot submit your samples equally 'premixed', ensure that the primer is mixed with your purified/prepared template. Afterward assembling the Templates and Primers as described in the tabular array ane and prepare your sample past mixing the following volumes of the two stock solutions.
Tabular array ii: Assembling Premixed samples
| Component | Volume (μL) |
| Template DNA | 5 |
| Primer | 5 |
| Total (Sample) | 10 |
Note that only the premixed samples must be submitted for ShortSeq plate and the SimpleSeq Premixed sequencing services. Ensure that the samples for 'ShortSeq plates' or 'SimpleSeq Kits - Premixed' are prepared based on the guidelines prepare up here for 'Premixed' samples.
'Templates Only' Samples
When you practise not have the standard or custom primers handy or ready for apply, you can submit 'Templates Merely' Samples. For the primer required in this sequencing reaction, you lot can
- Use one of our >75 universal primers to be added to your sample earlier sequencing (free), or
- Society the synthesis of a custom primer that volition then be used for sequencing your samples and subsequently stored in our facility for your future sequencing orders
Table three: Assembling 'Templates Only' Samples.
| Sequencing chemistry | Template Stock solution — Book (μL) | |
| First Reaction | Each additional reaction | |
| Standard Read | 10 | 5 |
| Ability Read | 10 | 10 |
'Templates and Primers' Samples:
When you submit 'Templates and Primers' Samples, our sequencing technicians will mix the primer(s) with your sample(s) prior to sequencing. The power to send primers and templates separately allows you to submit samples for multiple sequencing reactions or society additional services such as PCR purification/DNA Preparation for your template sample. The following table provides guidelines for the submission of prepared templates and primers. Use the concentration of templates and primers in tabular array 1 to gear up the stock solutions. When you submit 'Templates and Primers' samples:
- You must submit your primers and templates in separate containers,
- Concentrations of template Deoxyribonucleic acid and Primers must adhere to the recommended values (come across table ane).
- Provide sufficient quantities of samples and primers (see table below). You can order multiple reactions for a template submitted in one tube or a unmarried well of the 96-well plate. Please annotation that the volume requirements vary for standard and power read sequencing services.
- Use barcode labels (optional) on the primer and sample containers,
- Indicated on the tube sequencing (cheque box) or plate sequencing (mention in notes) order editor if you want the sent primer to be stored at our facility for your future orders. On request, these primers will be stored in our facility for 1 month or till they are wearied, which e'er is earlier.
Table 4: Assembling 'Templates and Primers' samples
| Sequencing chemistry | Template Stock solution — Volume (μL) | Primer Stock Solution — Volume (μL) | ||
| First Reaction | Each additional reaction | First Reaction | Each additional reaction | |
| Standard Read | 10 | v | ten | 5 |
| Power Read | 12 | 12 | 12 | half dozen |
Samples Requiring 'Boosted Services':
When your sample requires further preparation or purification prior to sequencing, the post-obit atmospheric condition should be met:
- The template(s)/Sample(s) and primers(when enclosed) should exist submitted in separate containers
- The volume of the sample based on its blazon should see the following requirements (see table).
Tabular array 5: Assembling samples which require additional services
| Sample Blazon | Type of service | Sample Solution — Volume (μL) | [Primer] (pmol/μL) | Primer(s) stock solution — Volume (μL) |
| Crude PCR Products (>100 bp) | PCR Purification | 15–thirty (Unabridged Reaction Volume) | ii–10 | x (5 for each improver rxn) |
| Bacterial Cultures | RCA Plasmid training | 10 (Glycerol stock recommended) | 2–10 | 10 (5 for each add-on rxn) |
Preparation of 'Ready2Load' Samples
Ready2Load sequencing services is provided for samples that have a single requirement: to be run on the sequencing gel. Samples beingness submitted as Ready2Load samples must meet the following requirements:
- The BigDye v3.ane reaction must be completed.
- The purification steps that remove unincorporated dye molecules must have been completed.
- It is recommended that these samples are sent every bit dry pellets.
Note that the Ready2Load plates are processed using an abbreviated protocol which typically take a read length of 500–700 bp.
Storage
Information almost Eurofins' storage policy is captured and categorized in the sections above.
For customers who need to store their sample(s) before submitting it, nosotros recommend storing both samples and primers in a freezer.
Additional Links
Types of Services and accustomed Samples
We have tailored our Deoxyribonucleic acid sequencing services using the Sanger method considering the diverse natures of the projects and requirements of the researchers. Researchers tin can transport different types of samples in unlike formats with or with out including the primers or requiring additional services on the samples.
| Sample/Services Feature | Description |
| Services Provided | The post-obit services are provided:
|
| Accepted Sample types | The post-obit types of templates are accepted:
|
| Sequencing Chemistries | Standard Read - Suitable for nigh standard templates requiring read lengths of upto 700 bps Power Read - Suitable for templates with difficult to read segments in the sequence. These tin include:
|
| Accustomed Sample containers | These include:
|
Sequencing Services provided for various Sample Types based on the inclusion of Sequencing primers
Each of the sequencing reaction requires a Sequencing primer that is specific to the template being read. Nosotros provide the most widely used sequencing primers for no additional cost. Notwithstanding, if you practice need custom primers, our DNA synthesis lab will synthesize and deliver the aforementioned to the sequencing lab with remarkable turnaround times.
| Sample Types | Description |
| Premixed | Submit the template of interest premixed with sequencing primer to receive the results from a standard read in the shortest time. |
| Templates Merely | Submit but your template. You can choose from a broad range of services for samples submitted as templates only. These include:
|
| Templates and Primers | When you send your sample templates and primers in dissever containers, we can provide the following additional services:
|
| Custom | Research Services: Whether yous demand the sequence of a large construct such as BACs or Fosmids, rough PCR product, or you just need the sequencing gel run afterwards having performed the BigDye® reaction, we take a service tailored for your need. If you practice not detect something for your specific need, please contact your sales representative or write to united states at genomicsUS@eurofins.com. Clinical Inquiry Services: Our CLIA and CAP accredited sequencing lab provides services suited to your specific needs. Contact our CLIA specialist for more information. |
#For more details on Next Generation Sequencing Services, click hither.
*Additional charges apply.
Boosted Links
| Tube Format <48 samples Submit Tubes All tube types are submitted on the same webpage except SimpleSeq Premixed which does not require online submission. |  | Barcoded Tubes
|
 | Non-Barcoded Tubes
| |
 | 8 Strip PCR Tubes
| |
 | SimpleSeq Tubes
|
| Plate Format >48 samples Submit Standard and Prepaid Plates Order Prepaid Plates Submit ShortSeq Submit Ready2Load |  | Sequencing Standard 96-well Plate |
 | Prepaid Plate Kit | |
 | ShortSeq Plate
| |
| | Ready2Load Plate
| |
Boosted Links
Download results
Go to your Order History. Next, click the gild to expand and reveal the download link. Click the link to download a aught file of the results. To visit the Order History folio, click i of the many links located throughout the website, including in the header beside your account proper name.
When looking at the order history folio, each blazon of order is color coded by its production line, so it is easy to discover the order you want. For example, sequencing orders are marked by a modest blue icon, oligo orders are marked by an orange icon, and so forth. Delight note that sequencing results are only saved for a limited amount of time. If you are trying to download very erstwhile results, it may not exist available online merely may be archived and bachelor by manual retrieval. Contact us for more information.
Guidelines for Overnight Sequencing Service
Overnight Sequencing service from Eurofins Genomics allows you to advance your research project in an uninterrupted manner inside a limited budget. To avail the Overnight Sequencing service from Eurofins Genomics, your society/samples must meet the following requirements:
- The samples are shipped using 'Overnight Aircraft' service
- The social club must require less than 192 sequencing reactions
- This services applies merely to samples requiring 'Standard Read' sequencing chemistry (this does non include other services such as power reads, BAC, PCR purification or RCA)
- Samples must exist supplied in i of the following containers
- 1.5-mL or two-mL centrifuge tubes with barcode labels* pasted on them
- 96-well plates with a gratis barcode label pasted at the bottom of the plate
- Use barcode-labeled tubes or semi-skirt Plates supplied by Eurofins Genomics, east.g., every bit tubes from SimpleSeq Kits or 96-well plates from the PrePaid plate Kits
- Use of Spiral Caps or Sealing using parafilms prevents cross contamination, only can extend the turnaround time.
Additional Links
Source: https://eurofinsgenomics.com/en/products/dna-sequencing/seq-sample-submission-guidelines/
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