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

(Wednesday 5th August)
(Ligation)
Line 24: Line 24:
 
Biobricks:  
 
Biobricks:  
 
* BBa_K1707020 : BBa_K1707004 + BBa_K1707019
 
* 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 Tampon 10x
+
** 2 µL Tampon 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 Tampon 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 Tampon 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 Tampon 10x
* 1 µL Ligase
+
** 1 µL Ligase
* 0,75 µL H2O
+
** 0,75 µL H2O
 
+
  
 
===Rehydratation===
 
===Rehydratation===

Revision as of 09:38, 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

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

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