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

@@ Line 1: / Line 1: @@
+{{Team:Paris_Saclay/notebook_header}}
 =Wednesday 5th August=
 ==Lab Work==
-===Plasmid extraction===
+===Quantification===
-''by Pauline''
+''by Pauline and Coralie''
-Biobricks:
+* BBa_R0051
-* BBa_K1707008 #1 to #6
+* BBa_E0022 (15ng/mL)
-* BBa_I13600 #1 and #2
+* BBa_E0422 (20ng/mL)
+* BBa_K1707000 (20ng/mL)
+* BBa_K1707004 (15ng/mL)
+* BBa_K1707011
+* BBa_K1707012
+* BBa_K1707013 (20ng/mL)
+* BBa_K1707019 (15ng/mL)
-With the Macherey-Nagel Extraction kit
+Agarose gel, 1%
+Migration 90V
-===Digestion Verification===
-''by Pauline and Coralie''
-Biobricks: BBa_K1707008 #1 to #6
+===Ligation===
+''by Pauline''
+BBa_K1707020:
+* BBa_K1707004 + BBa_K1707019
 Mix:
-* 2 µL Plasmid
+* 6,5 µL BBa_K1707019
-* 0,5 µL  XbaI
+* 9,5 µL BBa_K1707004
-* 0,5 µL PstI
+* 2 µL Buffer 10x
-* 1 µL Buffer FastDigest 10x
+* 1 µL Ligase
-* 6 µL H2O
+* 1 µL H2O
-Incubation 2h, 37°C
+BBa Test:
+* BBa_K1707004 + BBa_K1707013
+Mix:
+* 5 µL BBa_K1707013
+* 7,5 µL BBa_K1707004
+* 1,5 µL Buffer 10x
+* 1 µL Ligase
-===Electrophoresis===
-''by Pauline''
-Agarose gel 1%
+BBa_K1707027:
-Migration 90V
+* 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
-We can conclude that all BBa_K1707008 clones are OK
-We make a glycerol stock of #1 and #2
+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
-===Electrophoresis===
+===Rehydratation===
-''by Pauline and Coralie''
+''by Audrey''
-Agarose gel, 1%
+* BBa_R0040
-Migration 100V
+===Transformation===
+''by Audrey and Coralie''
-We can conclude that:
+* BBa_R0040
-* BBa_K1707021: #2, #3, #4 and #6 are OK
+* BBa Test
-* BBa_K1707011: #1 to #6 are OK
+* BBa_K1707028
-* BBa_K1707004: #1 to #6 are OK
+* BBa_K1707027
+* BBa_K1707020
-We make glycerol stock of them.
 ===Digestion===
-''by Coralie''
+''by Pauline''
 XbaI + PstI:
+* BBa_K1707012
 * BBa_K1707011
-* BBa_K1707004 x2
+* BBa_I13600
-* BBa_E0022
-* BBa_E0422
-* BBa_K1707012
-SpeI + Pst
+SpeI + Pst:
-=== New culture===
+* BBa_R0051
-'' by Pauline''
-Biobricks:
 * BBa_K1707008
-* BBa_I13600 (2014)
-clones of each in 5mL LB + 20µg/mL of chloramphenicol
+EcoRI + SpeI:
+* BBa_K1707004
-Incubation at 37°C, 250 rpm
+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''
-====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
-===Digestion===
+===Low Budget Challenge===
-''by Audrey and Coralie''
+''by Coralie''
-Biobricks:
-* BBa_K1707011 #1 to #6
-* BBa_K1707021 #1 to #6
-* BBa_K1707004 #1 to #6
-Mix:
-* 2 µL Plasmid
-* 1 µL Buffer FastDigest 10x
-* 0,5 µL XbaI
-* 0,5 µL PstI
-* 6 µL H2O
+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:'''

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

Latest revision as of 07:34, 15 September 2015

Contents

Wednesday 5th August

Lab Work

Quantification

Ligation

Rehydratation

Transformation

Digestion

Soil experiment

Water

Low Budget Challenge