Please find below the sample preparation instructions to perform your sequencing in an optimal way with GATC Biotech

• Plasmid DNA
• PCR fragments

Concentration:

• Plasmids: 30-100 ng/µL
• PCR fragments: 10-50 ng/µL
• Custom primers: 10 pmol/µL

Volume:

• DNA samples & primers: 20 µL each (sufficient for 8 SUPREMERUN tube reactions).
• DNA and primers to be delivered en 1.5 mL reaction tubes each.

Primer characteristics

The melting temperature (TM) of the primer should be between 52°and 58°C and the length should be between 17-19bp. Ideally, the GC content should be 10 G+C for a 17mer; 8-9G+C for an 18mer and 7-9 G+C for a 19mer. G or C should be at the 3’ end, but not more than 3 Gs or Cs. The primer sequence should be a good mix of all 4 nucleotides with no more than 4 identical bases in a row (AAAA or GGGG).

List of GTAC free universal primers

Storage of DNA and primer

Storage of DNA and primers delivered by customer: 10 days Custom primers synthesized through GATC Biotech will be stored for 6 months. For a new order after these 6 months of storage and within the following 6 months, a new primer aliquot will be ordered free of charge.

Delivery time

• Next business day following sample receipt 

• Purified plasmid DNA or purified PCR fragments with primer premixed in tubes

Purified plasmid DNA or purified PCR fragments with primer premixed in tubes Premix template DNA of 5 µl with either of following concentration:

• Purified plasmid DNA: 80 - 100 ng/µl
• Purified PCR product: 20 – 80 ng/µl

Add 5 µl of primer with following concentration:

• 5 µM (5 pmol/µl)

Please send total sample amount of 10 µl in 1,5 ml tubes

To ensure the best possible quality of sequence data, the total volume of the sample should not be less than 10 µl. We recommend measuring the DNA concentration on an agarose gel. Sequencing reactions cannot be repeated and all reactions will be charged for. Your DNA material cannot be stored.

• next business day upon sample receipt

If you have any questions concerning your results please contact our support to discuss about your results and we will do our best to improve them

• by phone : 09 70 44 67 43
• by e-mail : a.kohnke@gatc-biotech.com

You will be supported in French, English and German