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

Prasad Reddy, Keith McKenney
1996 BioTechniques  
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 10 12 phage particles/mL in 3 hours if the cells 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 × 10 7 cells) and still yields about 10 12 phage
more » ... phage particles/mL. These amounts are sufficient to purify 10-15 µ g of ssDNA and to carry out at least 10-15 DNA sequencing reactions. Bldg. 222, Rm. A 359, Biotechnology Division,
doi:10.2144/96205st05 pmid:8723931 fatcat:jg66sxxxpbfobnglmqhmjobkaq