Team:Paris Saclay/Notebook/August/6
Contents
Thursday 6th August
Lab Work
Soil experiment
by Coralie
Water
Observation of plates (05/08/2015) Almost all plates are yellow. We choose to stop the experiment.
We will define a new protocol:
- Count of bacteria before contamination
- Evaluation of the count of bacteria we should see at Day0
- Measurement of the pH before each sampling (We think that maybe the water pH change with adding the LB)
Low Budget Challenge
by Coralie
We can't say a lot of things... But we can see some colony on the agar with the baker's yeast in the agar. We try another time with different agar recipes:
- Agar 1:
- 0,27g Stock Cube
- 0,600g Agar Agar
- 0,135g Baker's yeast
- 0,225g sugar
- 45mL mineral water
- Agar 2:
- 0,27g Stock Cube
- 0,600g Agar Agar
- 0,135g Baker's yeast
- 45mL mineral water
We make striations with BBa_J23101+BBa_I13504 strain with a curved pin. We do the same on an usual LB plate without antibiotics (Control +)
Incubation over night, 37°C
Purification agarose gel
by Pauline and Audrey
- BBa_K1707012
- BBa_K1707004
- BBa_K1707011
- BBa_R0051
- BBa_K1707008
- BBa_I13602
- BBa_I13600
And PIJ773 digested by NotI
Agarose gel 1%
Migration 90 V
Verification of gel purification, from left to right: 1. DNA Ladder, 2. BBa_K1707011, 3. BBa_K1707004, 4. BBa_K1707004, 5. BBa_E0022, 6. BBa_E0422, 7. BBa_K1707012, 8. BBa_I13602, 9. Empty, 10. Empty
We made a mistake: BBa_K1707008, BBa_I13602 and Bba_R0051 shouldn't be purified on agarose gel. We can observe a big smear for PIJ773. We can also observe that BBa_I13602 has three bands. Rest is alright.Purification
by Pauline
- BBa_K1707011 #1
- BBa_K1707004 #1
- BBa_K1707012 #1
- BBa_R0051
- BBa_K1707008 #1
- BBa_I136000
- BBa_I13602
With Macherey-Nagel Kit
Quantification
by Pauline and Audrey
- BBa_K1707011 #1
- BBa_K1707004 #1
- BBa_K1707012 #1
- BBa_R0051
- BBa_K1707008 #1
- BBa_I136000
- BBa_I13602
PIJ773 digested by NotI
Agarose gel, 1%
Migration 100V
Well 1: Verification by digestion with NotI, Wells 3-9: Verification of gel purification; from left to right: 1. BBa_PIJ773, 2. DNA Ladder, 3. BBa_K1707004#1, 4. BBa_K1707008#1, 5. BBa_K1707011#1, 6. BBa_K1707012#1, 7. BBa_R0051, 8. BBa_I13600, 9. BBa_I13602, 10. Empty
Ligation
by Audrey
BBa_K1707026:
- BBa_E0422 + BBa_J23101
Mix:
- 11 µL BBa_E0422
- 8 µL BBa_J23101
- 1,5 µL Buffer 10x
- 2 µL Ligase
- 2,5 µL H2O
BBa_K1707032:
- BBa_I13600 + BBa_K1707008
Mix:
- 2 µL BBa_K1707008
- 5 µL BBa_I13600
- 1 µL Buffer 10x
- 2 µL Ligase
BBa_K1707033:
- BBa_J23101 + BBa_K1707006
Mix:
- 6,5 µL BBa_K1707006
- 1,5 µL BBa_J23101
- 1,5 µL Tampon 10x
- 2 µL Ligase
- 3,5 µL H2O
Incubation ON, 4°C
New culture
by Pauline
- BBa_K1707020
- Plamsid test
- BBa_K1707027
- BBa_R0040
- BBa_K1707028
2clones in 5mL LB + antibiotics
Member present:
- Instructors: Claire
- Students: Coralie, Audrey and Pauline
eigem.parissaclay AT gmail DOT com
t@iGEMParisSaclay
aUniversité Paris Sud
a91405 Orsay, France
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