Difference between revisions of "Team:Paris Saclay/Notebook/August/5"

(Soil experiment)
(Soil experiment)
Line 114: Line 114:
 
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:
  
====Soil====
+
* MacConkey + Spectinomycin
Observation of plates (04/08/2015): 
+
* MacConkey + Tetracyclin
 +
* MacConkey + Chloramphenicol
  
* Soil + H2O on new MCK:  lots of colonies are visible
+
And only MacConkey for the Water + LB
* Soil + LB on new MCK: lots of colonies are visible
+
* LB on new MCK: nothing
+
* MCK alone: nothing
+
  
We can suppose two different hypothesis:
+
Incubation ON, 37°C
* 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.
+
===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
  
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:'''
 
'''Member present:'''

Revision as of 09:48, 10 August 2015

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

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

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