Improved Method for the Production of M13 Phage and Single-Stranded DNA for DNA Sequencing

Abstract

An improved method is described for the efficient production of M13 phage and M13 single-stranded (ss)DNA in a relatively short time period. Infection of E. coli (F′) cells with as few as 5 phage particles can yield 1012 phage particles/mL in 3 hours if the cells are grown in LB broth or SOB broth supplemented with about 5 mM Mg2+. The method tolerates large variations in the initial multiplicity of infection (5–5000 phage per 5 × 107 cells) and still yields about 1012 phage particles/mL. These amounts are sufficient to purify 10–15mg of ssDNA and to carry out at least 10–15 DNA sequencing reactions.