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

Biotechniques. 1996 Jun;20(6):1022-7. doi: 10.2144/96206st03.


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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.
  • Technical Report

MeSH terms

  • Bacteriophage M13 / genetics*
  • DNA, Viral / chemistry
  • Genetic Vectors / biosynthesis
  • Genetic Vectors / isolation & purification
  • Polyethylene Glycols / metabolism
  • Polymerase Chain Reaction
  • Sequence Analysis, DNA / methods*
  • Templates, Genetic*


  • DNA, Viral
  • Polyethylene Glycols