Design, Order & Manage Your Sequencing Primers

Sanger Sequencing Primers

Design, Order & Manage Your Primers for Sanger Sequencing

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

Your options at a glance:

Select from 80 standard primers on stock

Select and manage your favourite standard primers from more than 80 different validated standard primers. All standard primers are pre-defined, validated and free of charge.

Design your specific sequencing primers

With our sequencing primer design tool you can define specific primer sequences yourself. Optimal design criteria are applied to ensure the best possible specificity. With the "Order now" button you can add your primers directly to your DNA sequencing order.

Enclose your own sequencing primers

You can send us your sequencing primers in separate tubes together with your samples. Use our free primer barcodes for labelling. Please consider the optimal primer conditions and the specifications for concentration and volume in our Sample Sumbission Guides.

Order the synthesis of your sequencing primers

There is no reason to order your sequencing primer separately or elsewhere. Order the synthesis of the required primers directly when entering your sequencing order. We keep these primers for you for 4 weeks free of charge. Storage of synthesized primers for one year is possible at an additional cost. During this period, you can conveniently reuse them at any time.

Premix primer with DNA template

Sequencing primers can be added directly to your DNA template and sent to us as premixed sample with the TubeSeq Supreme Service, Mix2Seq Service, LightRun Services and the PlateSeq Service senden.

If you prefer to store your sequencing primers in your lab, just order our SeqPrimer.

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Related Information

Optimum Primer Conditions

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 ]

Literature

Standard Primer