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- You select and use one of our standard primers free of charge (see standard primer list on the right side)
- You send your own primer (10 µM) in a separate tube along with your samples.
- You use the “premixed” option and add the primer to the sample yourself (final concentration of the primer: 4 µM).
- You order a specific sequencing primer. Just follow the design guidelines further below and order the primer together with your sequencing reactions by using the “order now” option.
- You let Microsynth take care of primer design. In this case, please provide all essential information and we will do the design and subsequent synthesis.
Guidelines for Primer Design
Primer length should be about 20 bases (+/-2)
G/C content should be about 50 %
- Melting temperatures should be around 50°C
Avoid hairpins, palindromic sequence and dimers
Avoid primers with 90 % assembly with a second binding site or where the last 7 bases from the primer's 3'-end match perfectly with another site
Please remember that 30-60 nucleotides get lost between the 3'-end of the primer and the start of sequence. Take this into account when you choose your primer position
Your specific sequencing primers will be kept at our sequencing lab for at least 6 months (or for 12 months in case you have added them to your “Custom Primer List”). During this time these primers or aliquots thereof will be sent to you upon request.