TubeSeq Supreme

Your benefits at a glance using our TubeSeq Supreme or our TubeSeq NightXpress Sanger sequencing services:

• High quality read lengths of up to 1,100 bases
• Free Q20, Q30 or Q40 monitored single strand data available
• Advanced sequence analysis options (e.g. "N" for unclear bases) are available, for free
• Selectable file types and customised file names can be specified in your personal preferences
• Secure online archive and access to your sequence data 
• Repetition in case of a technical failure* included
• Free storage of template DNA, custom synthesised and enclosed primers - see "Material & Data Storage"
• Extended storage of ordered and enclosed primers on request 
• Easy re-order options from stored DNA samples

* A technical failure is defined as a chemical or technical (IT, machines) defect in our lab. In order to ensure that the sequencing quality is not affected by a technical failure, an internal quality control is performed with each sequencing run. In addition to that, a sophisticated software is analysing each plate considering parameters like quality values (QV) and reading lengths.

If you see "DONE_QV_NOT_MET" as a result of your sequencing reaction, it means that the internal quality control has passed, but some sample quality values have not been met. (e.g. sample or primer concentration or volume, sequence design or structure etc.).

Use the following concentration and volume for your samples:

* Primer must have a total volume of 20 µl (double distilled water or 5mM Tris-HCl); 2 µl of primer volume is required for every additional sequencing reaction.

Quantify the template concentration via agarose gel or a photometer to ensure accurate results.

Premixed samples (a mixture of template and primer):

• Templates should consist of 15 µl purified DNA with either of the concentrations given in the table above
• Add 2 µl of primer with a concentration of 10 pmol/µl (10 µM)
• Ensure that the total volume of your premixed sample is 17 µl

 
Please send your sample tubes according to the requirements below:

• Use 1.5 ml safe-lock tubes for your templates and primers
• Do not tape or wrap tubes with parafilm. Safe-lock tubes offer perfect sealing and evaporation protection
• Label your sample tube with one of your barcode labels. Affix the label horizontally
• Label your enclosed primers with our free Primer Barcodes

 
Use the following concentration and volume for your samples:

* Primer must have a min volume of 15 µl - 20 µl (double distilled water or 5mM Tris-HCl); 2 µl of primer volume is required for every additional sequencing reaction.

Quantify the template concentration via agarose gel or a photometer to ensure accurate results.

Please send your samples and primers according to the requirements below:

• Submit your purified DNA sample in one of your TubeSeq NXP tube
• Use 1.5 ml safe-lock tubes for your enclosed primers
• Do not tape or wrap the tubes with parafilm. 
• Label your enclosed primers with our free Primer Barcodes

Benefit from the greatest flexibility to handle your sequencing primers.

Thanks to our DNA synthesis lab, bioinformatic know how and internal IT solutions you can choose how to provide us with your sequencing primers:

• Select from standard primers on stock
  Select and manage your favourite standard primers from more than 80 different validated standard primers
• Order specific sequencing primers
  Specific primers can be ordered directly with your sequencing order. To benefit from the NightXpress service, the order must be submitted until 9:00 am CET. These primers are stored for 4 weeks and can be re-used during this time
• Enclose your own sequencing primers
  You can send us your sequencing primers in 1.5ml tubes along with your samples by using our Primer Barcodes. Enclosed primers are stored for 10 days.
• Premix primer with DNA template (for TubeSeq Supreme)
  Sequencing primers can be added directly to your DNA template and sent to us as premixed sample.

Please consider optimal primer conditions, concentration and volume as described below.

Optimum primer conditions:

• Primers must not contain phosphorylation or fluorescent dyes
• The optimum primer length is between 16-25 bases
• The primer melting temperature (Tm) should be 50 - 62°C
• The GC content of the primer should be 35-60%
• Ideally one G or C should be located at the 3' primer end
• The number of 3' Gs or Cs should not exceed 2 Gs or Cs
• If possible, avoid >3 identical bases in a row in the sequence

Primer concentration and volume:

• Exactly 10 pmol/µl primer concentration is required per sequencing reaction
• Each primer must have a total volume of 20 µl (double distilled water or 5mM Tris-HCl); 2 µl of primer volume is required for every additional sequencing reaction for the TubeSeq Supreme service. 5µl for the TubeSeq NightXpress service.
• Concentration of primers with wobble bases must be calculated according to the following formula: nX x ConcPrimer

n = number of bases within a wobble according to IUPC code, X = number of wobbles within the primer sequence. [e.g. 1 V (AGC) = 3¹ x 10 pmol/µl; 2 Y (CT) (CT) = 2² x 10 pmol/µl; 1 N (AGCT) = 4¹ x 10 pmol/µl ]

DNA and primer storage:

• Storage of template DNA for 4 weeks
• Storage of synthesised primers for 4 weeks free of charge
• Storage of synthesised primers for one year at additional cost
• Storage of enclosed primers for 10 days free of charge
• Storage of enclosed primers for 6 months at additional cost

Data storage:

• Secure online archive of all selected sequence data for 6 months

Benefit from a variety of online features to manage your prepaid barcode labels, TubeSeq NXP Kit and TubeSeq Coupons:

Multiple options to define the rights of use

• TubeSeq Labels and TubeSeq Supreme Barcodes can be used by anyone with the same customer number
• The usage rights can be restricted to your account in your sequencing preferences.
• You can also define a group of people who are allowed to use your TubeSeq Labels, TubeSeq Supreme Barcodes, TubeSeq Coupons and the tubes of your TubeSeq NXP Kit "My Groups".
• All type of prepaid tube barcodes can be transferred to any other account under "My Sanger Kits & Barcodes"
  
  

Automatic reminder to order again

• You always see the actual number of remaining prepaid reactions under "My Sanger Kits & Barcodes" where a traffic light signals when they are running low.
• In your sequencing preferences you can select to receive a reminder by email to order again when the critical minimum amount is achieved.
  
  

Overview of status and usage details

• Under the "Prepaid Tube Barcodes" tab on the "My Sanger Kits & Barcodes" page you can find full details of your purchased prepaid barcodes and Coupons.
• You can search, filter and sort to find specific barcodes.
• The details of users and sequencing orders are displayed in a tooltip by positioning your mouse cursor over the row of interest.

All prepaid barcode labels, kits and coupons are valid for 5 years and only expire after 5 years.

Important information:

Specified delivery times are mainly depending on sample arrival time in the lab.

For many European cities we are providing coordinated pick-up and transport services using Eurofins DropBoxes ensuring on time delivery in the lab.

However, these logistics services may also be subject to force majeure or unpredictable changes caused by e.g. strikes, accidents, storms, floods. In such events, Eurofins Genomics cannot guarantee the stated data delivery time and is not responsible for delays caused by logistics services.

Additionally, our general terms and conditions (GTCs) apply.