The quality and quantity of DNA template are two extremely critical factors for successful sequencing. A great variety of commercial kits are available for DNA template preparation. Some laboratory procedures (home-made protocols) have proven performance if you choose to do away from commercial kits.

For preparing plasmid (double-stranded) DNA templates, we strongly recommend the mini-prep Qiagen kits. Series of Qiawell and Qiaprep, whether in single tubes, 8-tube strips, or 96-well plates, have proven suitable for preparing small amount of samples for automated DNA sequencing. Qia Tip 20 is also effective and have the minimum requirement of lab equipment. Other commercial kits commonly used to prepare DNA templates for auto sequencing include Promega Wizard and Magic Miniprep Kits, 5'-3' Perfect Prep kit and Nucleobond AX cartridges. A general perception about these kits is that they may not produce consistent quality templates despite the fact that a number of labs have been using them with success. Detailed information on these preparation methods is available at the facility.

For manually preparing plasmid DNA for automated sequencing, PE Biosystems recommends a modified alkaline-lysis/PEG precipitation procedure ( See Appendix A for details).

For sequencing of single stranded M13 templates, ABI recommends the DynabeadsÆ lac Z Vector Purification Kit from Perkin-Elmer (P/N 401436).