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    TubeSeq Supreme

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    Sanger sequencing of plasmids and PCR products in tubes with TubeSeq Supreme

     

     

    TUBESEQ SupremE

    Prepaid Barcodes

    Primer Barcodes

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    Highlights of the TubeSeq Supreme:

     

    • All types of sample material, incl. large constructs are possible
    • PCR purification is available as additional service
    • Advanced options for complex sequence regions (extended cycle sequencing,  special chemistry) can be selected per reaction
    • Up to 8 reactions per sample can be ordered or re-ordered from stored samples
    • Flexible payment options with prepaid barcodes and coupons or on account
    • Data delivery next business day until 6 pm CET

    The benefits at a glance:

     

    • High quality read lengths of up to 1,100 bases
    • Free Q20, Q30 or Q40 monitored single strand data available
    • 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.).

     

     

     

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    Related Information

     

     

    Sample Preparation & Submission TubeSeq Supreme


    Use the following concentration and volume for your samples:

    Sample type
    Product length
    Sample conc.
    Vol. 1-8 rct.
    Plasmid DNA < 30 kbp 50-100 ng/µl 20 µl
    BAC/PAC/Cosmid DNA 30 - 200 kbp 200-1000 ng/µl 20 µl
    Purified PCR Products 150-300 bp 1 ng/µl 20 µl
      300-1000 bp 5 ng/µl 20 µl
      1000-3000 bp 10 ng/µl 20 µl
    Unpurified PCR Products 150-300 bp 4 ng/µl 20 µl
      300-1000 bp 10 ng/µl 20 µl
      1000-3000 bp 20 ng/µl 20 µl
    Enclosed Primer 16-25 bases 10 pmol/µl 20 µl*

    * 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

     

     

     

    Sequencing Primers


    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 ]

     

     

     

    Material & Data Storage

     

    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

     

    Prepaid Barcodes & Coupons


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

     


    Multiple options to define the rights of use

    • 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 Supreme Barcodes and TubeSeq Coupons  "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.

     

     

    Disclaimer for Data Delivery


    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.

     

     

    Literature

     

    TubeSeq Supreme sample submission guide

    Sequencing result guide

    Free standard primers

    DNA sequencing brochure

     

     

     

     

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    Our products and services are produced and performed under strict quality management and quality assurance systems.

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