Products will be sequenced if any
- redo of ones that gave 500/1kb bands from 11/30 cco KO's: colonies 1, 3, 8, 9, 10
- brown cco: 3 from supernatant and 1 from pellet
- brown cbb: 2 from supernatant and 1 from pellet
25ul reactions, 40 cycles:
- 7 min initial 94C denaturation
- 30 s denature
- 30 s annealing 58→56.5 in first 15 cycles by dropping 0.1 decrease/cycle, 59→56.5 in last 25 cycles
- 2 min elongation
- 10 min final extension
conjugation, round 2
- no MR-1 colonies yet on plates off filter
- maybe should supplement with ferric citrate?
- new LB/no salt cultures for new cco, cbb#3, and Duo#18, supplemented with 500ul of filtered ferric citrate solution (in 8mL salt free LB, concentration unknown...)
- can it/ need it be done under anaerobic conditions?