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

Prasad Reddy | Zendy; Keith McKenney | Zendy

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

Author(s) -

Prasad Reddy,

Keith McKenney

Publication year - 1996

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/96205st05

Subject(s) - dna , multiplicity of infection , phagemid , dna sequencing , microbiology and biotechnology , biology , phage display , bacteriophage , bacteria , dna synthesis , escherichia coli , chemistry , gene , genetics , antibody

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 Mg2+. 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 micrograms of ssDNA and to carry out at least 10-15 DNA sequencing reactions.

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