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Xylose test

  • Preparation of xylose tube

- Add 1mL of culture from yesterday, 1mL of xylose (0.1g/mL) and 8mL of LB and 45μL of Spc (do that for the 4 different colonies)

- Incubate 5hours

- Observation with microscope : some green task, but they do not really match with bacillus cells, not conclusive!

Starch plate

- Add 5mL of iodine solution on starch plate, wait for 1min, throw the liquid

- Result

Strain added vector Observation Conclusion
IA771 none no clearance zones ok
IA751 none big clearance zones ok
IA751 ECE153 no clearance zones trnsformation worked!
IA751 or IA771 none or ECE171 big clearance zones we have some colonues on control plate with IA751, contamination? or antibiotic resistance?

Ery test

- IA771 alive and IA751 dead, confirmation that we did not mix plates (same conclusion than with starch plate, last line)

Test glycerol stocks of competent bacillus

- Pellet bacillus cells, add 100μL of prewarmed mediumB, add 0.6μg of DNA, incubate 30min at 37°C by shaking, plate

Strain added vector Antibiotic
IA771 none blank plate (to check they are alive)
IA771 none Cm5 (negative control)
IA771 ECE166 Cm5
IA751 none blank plate (to check they are alive)
IA751 none Cm5 (negative control)
IA751 none Spc50 (negative control)
IA751 ECE112 Cm5
IA751 ECE153 Spc50