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

TitleMethod for 96-well M13 DNA template preparations for large-scale sequencing.
Publication TypeJournal Article
Year of Publication1996
AuthorsAndersson, B, Lu, J, Edwards, KE, Muzny, DM, Gibbs, RA
Date Published1996 Jun
KeywordsBacteriophage M13, DNA, Viral, Genetic Vectors, Polyethylene Glycols, Polymerase Chain Reaction, Sequence Analysis, DNA, Templates, Genetic

Efficient preparation of DNA templates is an important step in large-scale DNA sequencing. The ensure high-quality sequence data, we have prepared M13 phage DNA templates using a glass fiber-filtration method. We present the adaptation 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.

Alternate JournalBiotechniques
PubMed ID8780873
Grant ListP30 HG00210-05 / HG / NHGRI NIH HHS / United States
R01 HG00823-01A1 / HG / NHGRI NIH HHS / United States

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