Stored in 1% glycerol in same freezer as P. haloplanktis
Scrape a little bit of ice (Frozen Bacteria) with a tip and put it into 20 ml 1%TB
Incubate at 30 degrees at 250 rpm in shaker inside incubator in cabinet in Polz lab for minimum 14 hours OVERNIGHT (ON)
Take 50uL of the (ON) culture and put it into a new tube with 5 mL of 1%TB
Incubate at 35 degrees at 250 rpm until OD = 0.4-0.45 (should take approx. 5 hours 40 min)
Wash
- Centrifuge at 4500 rpm for 5 min
- Resuspend pellet in Howard Berg medium (suck out supernatant after centrifuging (900 ul) add 900 ul of HB medium
- Repeat x 2