title: Cost effective DNA sequencing and template preparation from bacterial colonies and plasmids.
abstract: Cost-effective DNA sequencing and template preparation were evaluated for high-throughput screening of bacterial colonies. The rolling-circle amplification to generate template for DNA sequencing was carried out using 10-fold smaller amount of phi29 DNA polymerase and excluding two other enzymes (yeast pyrophosphatase and calf intestine alkaline phosphatase) used previously. Then, a 1/40 volume of the fluorescent terminator mix recommended by the manufacturer gave a usable sequencing result although a 1/26.7 volume of the mix was used to compare the protocol more accurately with manufacturer's protocol.
authors.affiliation: Division of Bacteriology, International Research Center for Infectious Diseases, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo, Japan.
authors.forename: Fumito
authors.initials: F
authors.lastname: Maruyama
authors.forename: Takashi
authors.initials: T
authors.lastname: Nozawa
authors.forename: Chihiro
authors.initials: C
authors.lastname: Aikawa
authors.forename: Atsuo
authors.initials: A
authors.lastname: Sakurai
authors.forename: Ichiro
authors.initials: I
authors.lastname: Nakagawa