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

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

Research output: Contribution to journalArticle

15 Citations (Scopus)

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
StatePublished - 1996

Fingerprint

Bacteriophages
DNA
Bacteriophage M13
DNA Sequence Analysis
Data Accuracy
fiberglass
2-(2-(2-chloro-3-(2-(3,3-dimethyl-5-sulfo-1-(4-sulfo-butyl)-3H-indol-2-yl)-vinyl)-cyclohex-2-enylidene)-ethylidene)-3,3-dimethyl-1-(4-sulfo-butyl)-2,3-dihydro-1H-indole-5-carboxylic acid

All Science Journal Classification (ASJC) codes

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

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.
Andersson, B. ; Lu, Junxuan ; Edwards, K. E. ; Muzny, D. M. ; Gibbs, R. A. / Method for 96-well M13 DNA template preparations for large-scale sequencing. In: BioTechniques. 1996 ; Vol. 20, No. 6. pp. 1022-1027.
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Andersson, B, Lu, J, Edwards, KE, Muzny, DM & Gibbs, RA 1996, 'Method for 96-well M13 DNA template preparations for large-scale sequencing', BioTechniques, vol. 20, no. 6, pp. 1022-1027.

Method for 96-well M13 DNA template preparations for large-scale sequencing. / Andersson, B.; Lu, Junxuan; Edwards, K. E.; Muzny, D. M.; Gibbs, R. A.

In: BioTechniques, Vol. 20, No. 6, 1996, p. 1022-1027.

Research output: Contribution to journalArticle

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AU - Lu, Junxuan

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AB - 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.

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Andersson B, Lu J, Edwards KE, Muzny DM, Gibbs RA. Method for 96-well M13 DNA template preparations for large-scale sequencing. BioTechniques. 1996;20(6):1022-1027.