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

E W Skowronski; N Armstrong; G Andersen; M Macht; P M McCready

doi:10.2144/00294st05

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PubMed · 11056809

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

E W Skowronski·N Armstrong·G Andersen·M Macht·P M McCready

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Abstract

We have developed a rapid, microplate-format plasmid isolation procedure to purify sequencing-grade DNA templates for high-throughput DNA sequencing operations. A modified lysozyme/boiling method is used to produce a plasmid-containing supernatant that is then purified by iron bead capture. After binding, the beads are pelleted in a magnetic field, washed and the DNA eluted in water. The method yields up to 10 micrograms plasmid DNA from a 1-mL overnight culture in a deep-well microplate. The procedure is suitable for large-scale experiments, amenable to automation and does not require expensive reagents 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 up to 48 plates per day. This protocol is ideal for any high-throughput operation in which template quantity, quality and reproducibility are of primary importance.

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DNA Plasmids Sequence Analysis, DNA Journal Article Research Support, U.S. Gov't, Non-P.H.S.

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E W Skowronski, N Armstrong, G Andersen, M Macht, P M McCready. 2000. Magnetic, microplate-format plasmid isolation protocol for high-yield, sequencing-grade DNA.. https://doi.org/10.2144/00294st05

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