Difference between revisions of "Team:Paris Saclay/Notebook/August/5"
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+ | {{Team:Paris_Saclay/notebook_header}} | ||
=Wednesday 5th August= | =Wednesday 5th August= | ||
==Lab Work== | ==Lab Work== | ||
Line 4: | Line 5: | ||
''by Pauline and Coralie'' | ''by Pauline and Coralie'' | ||
− | |||
* BBa_R0051 | * BBa_R0051 | ||
* BBa_E0022 (15ng/mL) | * BBa_E0022 (15ng/mL) | ||
Line 22: | Line 22: | ||
''by Pauline'' | ''by Pauline'' | ||
− | + | BBa_K1707020: | |
− | + | * BBa_K1707004 + BBa_K1707019 | |
− | + | ||
Mix: | Mix: | ||
* 6,5 µL BBa_K1707019 | * 6,5 µL BBa_K1707019 | ||
* 9,5 µL BBa_K1707004 | * 9,5 µL BBa_K1707004 | ||
− | * 2 µL | + | * 2 µL Buffer 10x |
* 1 µL Ligase | * 1 µL Ligase | ||
* 1 µL H2O | * 1 µL H2O | ||
− | + | BBa Test: | |
− | BBa Test : BBa_K1707004 + BBa_K1707013 | + | * BBa_K1707004 + BBa_K1707013 |
− | + | ||
Mix: | Mix: | ||
* 5 µL BBa_K1707013 | * 5 µL BBa_K1707013 | ||
* 7,5 µL BBa_K1707004 | * 7,5 µL BBa_K1707004 | ||
− | * 1,5 µL | + | * 1,5 µL Buffer 10x |
* 1 µL Ligase | * 1 µL Ligase | ||
− | + | BBa_K1707027: | |
− | + | * BBa_K1707000 + BBa_E0022 | |
− | BBa_K1707027 : BBa_K1707000 + BBa_E0022 | + | |
− | + | ||
Mix: | Mix: | ||
* 5 µL BBa_K1707000 | * 5 µL BBa_K1707000 | ||
* 9 µL BBa_E0022 | * 9 µL BBa_E0022 | ||
− | * 1,5 µL | + | * 1,5 µL Buffer 10x |
* 1 µL Ligase | * 1 µL Ligase | ||
* 3,5 µL H2O | * 3,5 µL H2O | ||
− | + | BBa_K1707028: | |
− | + | * BBa_R0051 + BBa_E0422 | |
− | BBa_K1707028 : BBa_R0051 + BBa_E0422 | + | |
− | + | ||
Mix: | Mix: | ||
* 6,25 µL BBa_R0051 | * 6,25 µL BBa_R0051 | ||
* 5,5 µL BBa_E0422 | * 5,5 µL BBa_E0422 | ||
− | * 1,5 µL | + | * 1,5 µL Buffer 10x |
* 1 µL Ligase | * 1 µL Ligase | ||
* 0,75 µL H2O | * 0,75 µL H2O | ||
Line 69: | Line 63: | ||
''by Audrey'' | ''by Audrey'' | ||
− | BBa_R0040 | + | * BBa_R0040 |
===Transformation=== | ===Transformation=== | ||
Line 114: | Line 108: | ||
Sea water + 1696 (Dil:1): We can see that the MCK is all yellow. | Sea water + 1696 (Dil:1): We can see that the MCK is all yellow. | ||
− | J1: | + | J1: we take 100µL of each water and put it on plates with specific antibiotic: |
− | + | * MacConkey + Spectinomycin | |
− | + | * MacConkey + Tetracyclin | |
+ | * MacConkey + Chloramphenicol | ||
− | + | And only MacConkey for the Water + LB | |
− | + | ||
− | + | ||
− | + | ||
− | + | Incubation ON, 37°C | |
− | + | ||
− | + | ||
− | + | ===Low Budget Challenge=== | |
− | + | ''by Coralie'' | |
− | + | ||
− | + | We can't say anything about the agar: is it a bacterial mat or nothing is on the plate? | |
+ | We choose to incubate another time over night plates at 37°C | ||
'''Member present:''' | '''Member present:''' |
Latest revision as of 07:34, 15 September 2015
Contents
Wednesday 5th August
Lab Work
Quantification
by Pauline and Coralie
- 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
BBa_K1707020:
- BBa_K1707004 + BBa_K1707019
Mix:
- 6,5 µL BBa_K1707019
- 9,5 µL BBa_K1707004
- 2 µL Buffer 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 Buffer 10x
- 1 µL Ligase
BBa_K1707027:
- BBa_K1707000 + BBa_E0022
Mix:
- 5 µL BBa_K1707000
- 9 µL BBa_E0022
- 1,5 µL Buffer 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 Buffer 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
Water
Observation of plates (04/08/2015) We can see a lots of pink colony. Sea water + 1696 (Dil:1): We can see that the MCK is all yellow.
J1: we take 100µL of each water and put it on plates with specific antibiotic:
- MacConkey + Spectinomycin
- MacConkey + Tetracyclin
- MacConkey + Chloramphenicol
And only MacConkey for the Water + LB
Incubation ON, 37°C
Low Budget Challenge
by Coralie
We can't say anything about the agar: is it a bacterial mat or nothing is on the plate? We choose to incubate another time over night plates at 37°C
Member present:
- Instructors: Claire
- Students: Coralie, Audrey and Pauline