Cafeteria roenbergensis

Strains, materials and methods

Strain description

Kindly provided by C. O'Kelly.

Culture and media

Marine water plus bacteria.

Purification of mtDNA

Isolation of total cellular DNA and subsequent separation of the A+T-rich fraction through CsCl/bisbenzimide centrifugation.

Cloning

Random fragments were generated by nebulization, fractionated according to size (0.5-1.0 kbp; 1.0-3.0 kbp), and cloned in Bluescript KS+.
Performed by I. Plante (Unit).

Sequencing

Manual Sanger sequencing of single-stranded DNA, using ³⁵S label. One custom primer was used for primer walking. Assembly was performed using the Staden BAP program.
Performed by D. St-Louis (Unit).