Title | Method for 96-well M13 DNA template preparations for large-scale sequencing. |
Publication Type | Journal Article |
Year of Publication | 1996 |
Authors | Andersson, B, Lu, J, Edwards, KE, Muzny, DM, Gibbs, RA |
Journal | Biotechniques |
Volume | 20 |
Issue | 6 |
Pagination | 1022-7 |
Date Published | 1996 Jun |
ISSN | 0736-6205 |
Keywords | Bacteriophage M13, DNA, Viral, Genetic Vectors, Polyethylene Glycols, Polymerase Chain Reaction, Sequence Analysis, DNA, Templates, Genetic |
Abstract | 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. |
DOI | 10.2144/96206st03 |
Alternate Journal | Biotechniques |
PubMed ID | 8780873 |
Grant List | P30 HG00210-05 / HG / NHGRI NIH HHS / United States R01 HG00823-01A1 / HG / NHGRI NIH HHS / United States |
Method for 96-well M13 DNA template preparations for large-scale sequencing.
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