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

摘要

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 I0~(12) phage particles/mL in 3 hours if the celts are grown in LB broth or SOB -broth supplemented with about 5 mM Mg~(2+), The 'method tolerates large variations in the initial multiplicity of infection (5-5000 phage per 5 x 10~7 cells) and still yields about 10~(12) phage particles/mL. These amounts are sufficient to purify 10-15 μg of ssDNA and to carry out at least 10-15 DNA sequencing reactions.