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''
  
−
Biobricks:
 
 
* BBa_R0051
 
* BBa_R0051
 
* BBa_E0022 (15ng/mL)
 
* BBa_E0022 (15ng/mL)
Line 22: Line 22:
 
''by Pauline''
 
''by Pauline''
  
−
Biobricks:  
+
BBa_K1707020:
−
BBa_K1707020 : BBa_K1707004 + BBa_K1707019
+
* 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 Tampon 10x
+
* 2 µL Buffer 10x
−
** 1 µL Ligase
+
* 1 µL Ligase
−
** 1 µL H2O
+
* 1 µL H2O
  
−
BBa Test : BBa_K1707004 + BBa_K1707013
 
  
 +
BBa Test:
 +
* 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 Tampon 10x
+
* 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 Tampon 10x
+
* 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 Tampon 10x
+
* 1,5 µL Buffer 10x
−
** 1 µL Ligase
+
* 1 µL Ligase
−
** 0,75 µL H2O
+
* 0,75 µL H2O
  
 
===Rehydratation===
 
===Rehydratation===
 
''by Audrey''
 
''by Audrey''
  
−
BBa_R0040
+
* BBa_R0040
  
 
===Transformation===
 
===Transformation===
Line 89: Line 91:
 
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===
 
''by Coralie''
 
''by Coralie''
  
−
====Soil====
+
====Water====
−
Observation of plates (31/07/2015):
+
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.
  
−
* Soil + H2O on new MCK: lots of colonies are visible
+
J1: we take 100µL of each water and put it on plates with specific antibiotic:
−
* Soil + LB on new MCK: lots of colonies are visible
+
−
* LB on new MCK: nothing
+
−
* MCK alone: nothing
+
  
−
We can suppose two different hypothesis:
+
* MacConkey + Spectinomycin
−
* The soil is contaminated
+
* MacConkey + Tetracyclin
−
* the MCK isn't OK for our experiment: now we add lactose in the preparation. We will try with sorbitol instead of lactose.
+
* MacConkey + Chloramphenicol
  
−
We try soil + H2O and soil +LB on:
+
And only MacConkey for the Water + LB
−
* MCK + Sorbitol: Without antibiotics, with Spectinomycine, Tetramycine or Chloramphenicol
+
−
* MCK + Lactose: Without antibiotics, with Spectinomycine, Tetramycine or Chloramphenicol
+
  
 
Incubation ON, 37°C
 
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

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

Back to the calendar

Contact
eigem.parissaclay AT gmail DOT com
t@iGEMParisSaclay
aUniversité Paris Sud
a91405 Orsay, France Copyright © 2015 iGEM Paris Saclay. All rights reserved All inspirations encouraged.
*This wiki was conceived during long nights of work and a great coffee experience in the beautiful Orsay, France.