Difference between revisions of "Team:Paris Saclay/Notebook/August/5"
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Line 97: | Line 97: | ||
XbaI + EcoRI: | XbaI + EcoRI: | ||
* BBa_I13602 | * BBa_I13602 | ||
+ | |||
+ | |||
+ | Mix: | ||
+ | * 10 µL Plasmide | ||
+ | * 1 µL Enzyme 1 | ||
+ | * 1 µL Enzyme 2 | ||
+ | * 2 µL Buffer FastDigest 10x | ||
+ | * 6 µL H2O | ||
===Soil experiment=== | ===Soil experiment=== |
Revision as of 09:42, 10 August 2015
Contents
Wednesday 5th August
Lab Work
Quantification
by Pauline and Coralie
Biobricks:
- BBa_R0051
- BBa_E0022 (15ng/mL)
- BBa_E0422 (20ng/mL)
- BBa_K1707000 (20ng/mL)
- BBa_K1707004 (15ng/mL)
- BBa_K1707011
- BBa_K1707012
- BBa_K1707013 (20ng/mL)
- BBa_K1707019 (15ng/mL)
Agarose gel, 1% Migration 90V
Ligation
by Pauline
Biobricks: BBa_K1707020 : BBa_K1707004 + BBa_K1707019
Mix:
- 6,5 µL BBa_K1707019
- 9,5 µL BBa_K1707004
- 2 µL Tampon 10x
- 1 µL Ligase
- 1 µL H2O
BBa Test : BBa_K1707004 + BBa_K1707013
Mix:
- 5 µL BBa_K1707013
- 7,5 µL BBa_K1707004
- 1,5 µL Tampon 10x
- 1 µL Ligase
BBa_K1707027 : BBa_K1707000 + BBa_E0022
Mix:
- 5 µL BBa_K1707000
- 9 µL BBa_E0022
- 1,5 µL Tampon 10x
- 1 µL Ligase
- 3,5 µL H2O
BBa_K1707028 : BBa_R0051 + BBa_E0422
Mix:
- 6,25 µL BBa_R0051
- 5,5 µL BBa_E0422
- 1,5 µL Tampon 10x
- 1 µL Ligase
- 0,75 µL H2O
Rehydratation
by Audrey
BBa_R0040
Transformation
by Audrey and Coralie
- BBa_R0040
- BBa Test
- BBa_K1707028
- BBa_K1707027
- BBa_K1707020
Digestion
by Pauline
XbaI + PstI:
- BBa_K1707012
- BBa_K1707011
- BBa_I13600
SpeI + Pst:
- BBa_R0051
- BBa_K1707008
EcoRI + SpeI:
- BBa_K1707004
XbaI + EcoRI:
- BBa_I13602
Mix:
- 10 µL Plasmide
- 1 µL Enzyme 1
- 1 µL Enzyme 2
- 2 µL Buffer FastDigest 10x
- 6 µL H2O
Soil experiment
by Coralie
Soil
Observation of plates (31/07/2015):
- Soil + H2O on new MCK: lots of colonies are visible
- Soil + LB on new MCK: lots of colonies are visible
- LB on new MCK: nothing
- MCK alone: nothing
We can suppose two different hypothesis:
- The soil is contaminated
- the MCK isn't OK for our experiment: now we add lactose in the preparation. We will try with sorbitol instead of lactose.
We try soil + H2O and soil +LB on:
- MCK + Sorbitol: Without antibiotics, with Spectinomycine, Tetramycine or Chloramphenicol
- MCK + Lactose: Without antibiotics, with Spectinomycine, Tetramycine or Chloramphenicol
Incubation ON, 37°C
Member present:
- Instructors: Claire
- Students: Coralie, Audrey and Pauline