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

(Soil experiment)
 
(One intermediate revision by one other user not shown)
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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 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 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
Line 69: Line 63:
 
''by Audrey''
 
''by Audrey''
  
BBa_R0040
+
* BBa_R0040
  
 
===Transformation===
 
===Transformation===
Line 129: Line 123:
 
We can't say anything about the agar: is it a bacterial mat or nothing is on the plate?
 
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 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

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