Revision as of 11:05, 6 August 2015

Monday 3rd August

Lab Work

New culture

by Pauline

Biobricks:

BBa_K1707008
BBa_I13600 (2014)

6 clones of each in 5mL LB + 20µg/mL of chloramphenicol

Incubation at 37°C, 250 rpm

Plasmid extraction

by Seong-Koo

Biobricks:

BBa_K1707011 #1 to #6
BBa_K1707021 #1 to #6
BBa_K1707004 #1 to #6

Plasmid pIJ773

With the Macherey-Nagel Extraction kit

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

Back to the calendar

@@ Line 47: / Line 47: @@
-===Rehydratation===
-''by Pauline''
-Biobrick: BBa_I13600 (2014-3-19C and 2013-3-20C)
-===Transformation===
-''by Pauline''
-Biobricks:
-* BBa_I13600 (2013, 2014 and 2015)
-* BBa_K1707008
-As usual
-===Plasmid extraction===
-''by Seong-Koo''
-Biobricks:
-* BBa_E0422 #1 and #2
-* BBa_E0022 #1 and #2
-With the Macherey-Nagel Extraction kit
-===New culture===
-''by Pauline''
-Biobricks:
-* BBa_K1707004: amicillin
-* BBa_K1707011
-* BBa_K1707021
-clones of each in 5 mL LB
-===Digestion Verification===
-''by Coralie''
-* Biobricks:
-** BBa_K1707012 #1, #2, #3 and #4
-** BBa_K1707013 #1, #2, #3 and #4
-** BBa_K1707019 #1, #2, #3 and #4
-* Mix:
-** 1 µL Buffer FastDigest 10x
-** 0,5 µL XbaI
-** 0,5 µL PstI
-** 2 µL Plasmid
-** 6 µL H2O
-Incubation 2h, 37°C
-===Electrophoresis===
-''by Coralie''
-Agarose gel, 1%
-Migration 100V
-We can conclude that all biobricks are OK
-===Soil experiment===
-''by Coralie and Johan''
-====Soil====
-Observation of J0 plates: there are a lot of colony in each.
-Control (-):
-* MCK + Tet: 40 pink colonies
-* MCK + Cm: 10 pink colonies
-* MCK + Spectino: 1 pink colony
-* MCK: 100 pink colonies and a lot of white colonies
-We can suppose that we have a contamination between our experiment and control. Or maybe we have a MCK problem because we change it during the experiment.
-So, we decide to stop the experiment and do some test:
-* Soil + H2O on new MCK
-* Soil + LB on new MCK
-* LB on new MCK
-* MCK alone
-Incubation over night, 37°C
-=== New culture===
-'' by Pauline''
-Biobricks:
-* BBa_K1707005
-* BBa_I13602
-A plasmid: PIJ773
-Incubation at 37°C, 250 rpm
 '''Member present:'''
-* Instructors: Alice
+* Instructors: Claire
-* Students: Coralie, Audrey, Pauline, Johan and Seong-Koo
+* Students: Coralie, Audrey and Pauline
 {{Team:Paris_Saclay/notebook_footer}}

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