Difference between revisions of "Team:UPMC-Paris/Notebook"

Week 1 :

Competence of Bsu9716:

Bacteria from 2/8/16 in fridge (Dj)
50 ml LB and 10 uL Kan added (so total 150 mL and 15 uL Kan) and cells back in incubator
Spin (4000 rpm 10 min) 50 ml X 2 and used pellet for competency
Spinned again same way after pulling both pellet together
Changed medium (50 mL LB + 5 uL Kan) and put it back at 4 degrees

Stock solutions (Dj)

Name Mass (g) Total Volume (mL)
1.2% MgSO4 0.1023 12.5
10% (w/v) Bacto yeast extract 1.0096 10
1% (w/v) casamino acids 0.1039 10
0.1 M CaCl2 (110.98 g/mol) 0.1792 15

SpC medium made by Djamila
T base 20 ml
50% (w/v) filtered glucose 0.2 ml
1.2% (w/v) MgSO4 0.3 ml
10% (w/v) Bacto yeast extract 0.4 ml
1% (w/v) casamino acids 0.5 ml

ODi 0.5 and put back at 37 degrees with shaking
ODf 0.8

SpII medium made by Sofiane
T base 200 ml
50% (w/v) filtered glucose 2 ml
1.2% (w/v) MgSO4 14 ml
10% (w/v) Bacto yeast extract 2 ml
1% (w/v) casamino acids 2 ml
0.1 M CaCl2 1 ml

2 mL of SpC BSU in SpII and 90 min incubation 37 degrees with shaking
Note:
Originally 1.2% of the dihydrate form so made the calculations in function of that; and Sofiane had to make a second batch
50% (w/v) glucose was filtered
No tryptophan added for the cell growth

Plates from 03/08/2016 (Zinebe)
Contains lawn and colonies ⇒ Not sure which are the B. subtilis.
Diluted some of the lawn and the colony each in 200 uL LB and plated it on LB-Kan then 37 degrees overnight

Note:
LB used for dilution was contaminated
When picking the colony with a tip some of the lawn was scraped with it

Pick primers and autoclaved MilliQ at school

Plates (from 04/08/16) (DJ)
Both plates had lawns on them however the plate containing the colony (along with some lawn and labelled Isolate) was green which was weird

Took plate from 3/8/16 and streaked on a plate divided in 3 a single colony, the lawn, and for the last portion, 10 uL of the B subtilis solution in the fridge was diluted in 200 uL of LB before streaking it. The rest of B subtilis solution in the fridge was spinned, diluted in 10ml LB and divided in 2 x 50 mL tubes, 5 uL of Kan was added in each. Both were put back in the fridge

Tris HCl made by Kevin

All streaks grew well so we put them in fridge

Week 2 :

Ca me soule toujours autant

1. Benjamin : Il s'est fait chier
2. Nassim : Il a cassé les couilles
3. Sofiane : Il a tout fait cramer
4. Wilhelm : Il est reparti
5. Kévin : Il est pas venu
6. Perwin : Continuation de la quête chez les RG
7. Zinebe : Elle a volé le PLA
8. Djamila : Comme d'hab, le quota
9. Les autres : Vacances