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    SupremeRun Plate Service

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    • SupremeRun Plate

     

     

    High throughput sequencing of DNA samples in 96well plates.

     

     

      

    Primer Barcodes

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    Highlights of the SupremeRun 96:

     

    SupremeRun 96 is a high-throughput sequencing service for routine Sanger sequencing of 96 samples with the same primer on a plate.


    Automated workflows guarantee easy and convenient sample processing. Prepaid SupremeRun 96 barcodes enables transparent sample tracking during the automated sequencing workflows.

    • Sanger sequencing of plasmids or PCR fragments in plates
    • Reading lengths of up to 1100 Q20 bases 
    • Easy online ordering with simple invoicing

    Product details:

     

    • High-throughput Sanger sequencing of up to 96 samples on a microtiter plate
    • Full 96well microtiter plate sequencing for 96 samples with one primer 
    • Free use of a wide variety of standard primers
    • Sequencing reads automatically processed with superior data analysis software
    • Reads with up to 1100 Q20 bases
    • Optimised Sanger sequencing protocols for difficult templates (GC-rich sequences, bisulfit samples or low copy plasmids)
    • Free storage of DNA and primers
    • Repetition of sequencing reactions in case of technical failure*

     

     

     

    >> FAQ's Sanger Sequencing

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    Related Information

     

     

    Sample Preparation & Submission


    Sample preparation & sumbission for the SupremeRun 96 service

    • Please use fully skirted PCR plates  
    • Template concentration must be normalised across the plate
    • PCR product size should not vary by more than a factor of 3
    • Samples should be sent liquid in a total volume of 15 µl
    • Seal your plates using 8-cap strips to prevent material loss
    • Stick the SupremeRun 96 plate barcode on the plate edge
    • Label your enclosed primer with our free Primer Barcodes
    • Ship samples at ambient temperature to our sequencing lab

     

    Sample type
    Product length
    Sample conc.
    Sample vol.
    Plasmid DNA < 30 kbp 50-100 ng/µl 15 µl
    Purified PCR products 150-300 bp 1 ng/µl 15 µl
      300-1000 bp 5 ng/µl 15 µl
      1000-3000 bp 10 ng/µl 15 µl
    Unpurified PCR products 150-300 bp min. 4 ng/µl 15 µl
      300-1000 bp min. 10 ng/µl 15 µl
      1000-3000 bp min. 20 ng/µl 15 µl

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

     

    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. 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


    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 15 µl (double distilled water or 5mM Tris-HCl); 5 µl of primer volume is required for every additional sequencing reaction
    • 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 V (AGC) (AGC) = 32 x 10 pmol/µl]

     

    Storage Times


    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

     

    Data storage:

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

    Repetition in case of technical failure


    Definition of technical failure

    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 is run with each plate (for that purpose well H12 should be kept empty, otherwise a quality control is not possible). In addition to that, a sophisticated software is analysing each plate considering parameters like quality values (QV) and reading lengths. If these parameters fall below a certain percentage across the plate an additional manual check is performed. If a technical failure is confirmed by this manual check, the plate is getting re-sequenced.

     

     

    Literature

     

    SR 96 sample submission guide

    Sequencing result guide

    Free standard primers

     

     

     

     

     

     

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    SeqPrimer


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    PlateSeq Service


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    >> READ MORE

     

     

     

    PCR Primer


    The optimum primer for every
    PCR and real-time PCR assay


    >> READ MORE

     

     

     

    Mix2Seq


    The smartest sequencing solution for premixed samples


    >> READ MORE

     

     

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