Difference between revisions of "Team:Paris Saclay/Notebook/August/5"
(→Ligation) |
|||
Line 23: | Line 23: | ||
Biobricks: | Biobricks: | ||
− | + | BBa_K1707020 : BBa_K1707004 + BBa_K1707019 | |
+ | |||
Mix: | Mix: | ||
** 6,5 µL BBa_K1707019 | ** 6,5 µL BBa_K1707019 | ||
Line 31: | Line 32: | ||
** 1 µL H2O | ** 1 µL H2O | ||
− | + | BBa Test : BBa_K1707004 + BBa_K1707013 | |
+ | |||
Mix: | Mix: | ||
** 5 µL BBa_K1707013 | ** 5 µL BBa_K1707013 | ||
Line 38: | Line 40: | ||
** 1 µL Ligase | ** 1 µL Ligase | ||
− | + | BBa_K1707027 : BBa_K1707000 + BBa_E0022 | |
+ | |||
Mix: | Mix: | ||
** 5 µL BBa_K1707000 | ** 5 µL BBa_K1707000 | ||
Line 46: | Line 49: | ||
** 3,5 µL H2O | ** 3,5 µL H2O | ||
− | + | BBa_K1707028 : BBa_R0051 + BBa_E0422 | |
+ | |||
Mix: | Mix: | ||
** 6,25 µL BBa_R0051 | ** 6,25 µL BBa_R0051 |
Revision as of 09:39, 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
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