General Guidelines

The DNA template concentration should be 150-200 ng/ul, except for PCR products, for which the concentration should be 5-20 ng/ul. [Please be aware that it is important to be within the desired concentration range, particularly for sequencing of plasmids.]

Volume of DNA to be submitted should be at least 6ul for each reaction. If you must submit less than the requested volume (2.5ul is an absolute minimum), then write the submitted volume on the wall of the tube. When submitting samples in a microtiter plate, please include 8ul of excess template, e.g., 14ul per sample for sequencing with one primer and 20ul for two.

It is very important for quality sequencing that plasmid template concentration be accurate. For consistent results, plasmid template concentration should be estimated both by uv absorption and by gel electrophoresis with a standard. For PCR templates, it is usually sufficient to estimate concentrations from ethidium-stained gels with a standard. It is recommended that first-time users submit only a few samples.