Team:Paris Saclay/Notebook/July/31
Contents
Friday 31st July
Lab Work
Plasmid extraction
by Seong-Koo
Biobricks:
- BBa_K1707012 #1, #2, #3 and #4
- BBa_K1707013 #1, #2, #3 and #4
- BBa_K1707019 #1, #2, #3 and #4
With the Macherey-Nagel Extraction kit
Rehydratation
by Pauline
Biobrick: BBa_I13600 (2014-3-19C and 2013-3-20C)
Transformation
by Pauline
Biobricks:
- BBa_I13600 (2013, 2014 and 2015)
- BBa_K1707008
As usual
Plasmid extraction
by Seong-Koo
Biobricks:
- BBa_E0422 #1 and #2
- BBa_E0022 #1 and #2
With the Macherey-Nagel Extraction kit
New culture
by Pauline
Biobricks:
- BBa_K1707004: amicillin
- BBa_K1707011
- BBa_K1707021
6 clones of each in 5 mL LB
Digestion Verification
by Coralie
- Biobricks:
- BBa_K1707012 #1, #2, #3 and #4
- BBa_K1707013 #1, #2, #3 and #4
- BBa_K1707019 #1, #2, #3 and #4
- Mix:
- 1 µL Buffer FastDigest 10x
- 0,5 µL XbaI
- 0,5 µL PstI
- 2 µL Plasmid
- 6 µL H2O
Incubation 2h, 37°C
Electrophoresis
by Coralie
Agarose gel, 1% Migration 100V
We can conclude that all biobricks are OK
Soil experiment
by Coralie and Johan
Soil
Observation of J0 plates: there are a lot of colony in each.
Control (-):
- MCK + Tet: 40 pink colonies
- MCK + Cm: 10 pink colonies
- MCK + Spectino: 1 pink colony
- MCK: 100 pink colonies and a lot of white colonies
We can suppose that we have a contamination between our experiment and control. Or maybe we have a MCK problem because we change it during the experiment.
So, we decide to stop the experiment and do some test:
- Soil + H2O on new MCK
- Soil + LB on new MCK
- LB on new MCK
- MCK alone
Incubation over night, 37°C
New culture
by Pauline
Biobricks:
- BBa_K1707005
- BBa_I13602
A plasmid: PIJ773
Incubation at 37°C, 250 rpm
Member present:
- Instructors: Alice
- Students: Coralie, Audrey, Pauline, Johan and Seong-Koo