Isolation of mitochondria, CsCl/Hoechst dye gradient (by K.M. Lonergan in MWG's lab).
Cloning:
tRNA cluster 1-rnl-tRNA cluster 2-
rns-cox1/2: K.M. Lonergan (MWG's lab).
Entire genome: random fragments, generated by nebulization and
fractionated according to size (0.5-1.0 kbp; 1.0-3.0 kbp), cloned in
Bluescript KS+.
Sequencing strategy: Sanger sequencing of single-stranded DNA, using 35S label. Custom primers (13) used for limited primer walking. Mainly manual assembly, limited assembly using Staden's xbap.
Performed by: Sequencing of tRNA cluster 1-rnl-tRNA cluster 2-rns-cox1/2: K.M. Lonergan (MWG's lab). Cloning of complete genome: I. Plante (Unit). Sequencing of complete genome: I. Plante (Unit); rnl-rns region partially resequenced.