Method for 96-well M13 DNA template preparations for large-scale sequencing

B. Andersson, J. Lu, K. E. Edwards, D. M. Muzny, R. A. Gibbs

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

Efficient preparation of DNA templates is an important step in large- scale DNA sequencing. To ensure high-quality sequence data, we have prepared M13 phage DNA templates using a glass fiber-filtration method. We present the adaption of this protocol to a 96-well format using commercially available filter plates. Two variations are described: one using polyethylene glycol precipitation and a second where the phage particles are disrupted before filtration, thus eliminating the need for precipitation. Using either of these protocols, 96 templates can be prepared in less than 2 h. Sufficient DNA for 1-2 dye primer sequencing reactions is routinely obtained from 1 mL of culture, and the resulting sequence data are of high quality.

Original languageEnglish (US)
Pages (from-to)1022-1027
Number of pages6
JournalBioTechniques
Volume20
Issue number6
DOIs
StatePublished - Jan 1 1996

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

Fingerprint Dive into the research topics of 'Method for 96-well M13 DNA template preparations for large-scale sequencing'. Together they form a unique fingerprint.

  • Cite this

    Andersson, B., Lu, J., Edwards, K. E., Muzny, D. M., & Gibbs, R. A. (1996). Method for 96-well M13 DNA template preparations for large-scale sequencing. BioTechniques, 20(6), 1022-1027. https://doi.org/10.2144/96206st03