Magnetic, microplate-format plasmid isolation protocol for high-yield, sequencing-grade DNA

We have developed a rapid, microplate-format plasmid isolation procedure to purify sequencing-grade DNA templates for high-throughput DNA sequencing o perations. A modified lysozyme/boiling method is used to produce a plasmid- containing supernatant that is then purified by iron bead capture. After bi nding, the beads are pelleted in a magnetic field, washed and the DNA elute d in water. The method yields up to 10 mu g plasmid DNA from a I-mt overnig ht culture in a deep-well microplate. The procedure is suitable for large-s cale experiments, amenable to automation and does not require expensive rea gents or equipment. The entire protocol can be completed in as little as 2 h, and one technician with a 96-well pipetting station can process rip to 4 8 plates per day. This protocol is ideal for any high-throughput operation in which template quantity, quality and reproducibility are of primary impo rtance.