Sequenzierservice


Suitable Samples

  • Plasmid DNA from mini-, midi- or maxipreparations (column-purified or not)
  • PCR products (purified by gel electrophoresis, column, or enzymatically)

Purity of DNA

  • The DNA has to be free of proteins, RNA, or salts. Therefore, thoroughly wash all DNA samples with ethanol.
  • Dissolve DNA in water.
  • Determine the DNA concentration as precisely as possible, either photometrically or by comparing with mass standards on an agarose gel.

Required Amounts and Concentrations of DNA Per Sequencing Reaction

  • Plasmids: 0.3 – 0.5 µg in total at a concentration between 0.1 µg/µL and 0.5 µg/µL in water
  • PCR products: 1 – 2 ng per 100 bp in maximally 4 µL of water
  • Primers: 1 µL of a 4 – 5 µM solution in water; when using standard primers (see Excel file), simply specify the desired primer.
  • Please provide double amounts of all components to enable a second run of sequencing in case the first run fails.

Sequencing Orders

  • Please fill in the sequencing order form (Excel file) completely. When submitting pre-pipetted plates, please use this form and only use the plates provided by us.
  • Please restrict the names of DNA and primers to 10 characters each.
  • Samples can be delivered in room NC 6/166 between 8:00 and 18:00. Please place your samples in the marked compartment of the −20°C freezer and the sequencing order form in the tray on the windowsill. Outside the opening hours, please use the mailbox next to the door. Moreover, you can always deliver your samples to our mailboxes in ND 05 North or MA 01 North.

Contact

Sequenzierservice Biochemie
NC 6/166

Ruhr-Universität Bochum
44780 Bochum

Tel.: +49 234 32-23742
E-Mail: sequencer-biochemie@rub.de

Contact Persons

Björn PetersBjörn Peters
Room NC 6/166
Tel. +49 234 32-23742
E-Mail


Equipment

ABI3130xl
We use a capillary sequencer 3130xl Genetic Analyzer by Applied Biosystems.

Sample Preparation