Team:Paris Saclay/Notebook/July/2

From 2013.igem.org

(Difference between revisions)
(→detail)
 
(24 intermediate revisions not shown)
Line 1: Line 1:
{{Team:Paris_Saclay/incl_debut_generique}}
{{Team:Paris_Saclay/incl_debut_generique}}
 +
='''Notebook : July 2'''=
='''Notebook : July 2'''=
 +
=='''Lab work'''==
-
=='''summary'''==
+
==='''A - Aerobic/Anaerobic regulation system'''===
-
*Made a quantity control for fnr/PSB1C3 ligation products by using NanoDrop
+
[[File:PS_NP.jpg|right|250px]]
-
*performed 4 genetic transformations: LacZ(with plasmid PSB1A2) , AmilCP(with plasmid PSB1C3) and the products of ligation (fnr+plasmid PSB1C3) into competent cells. The 4th transformation was a control sample.
+
===='''Objective : obtaining BBa_K1155000'''====
-
*cloning. Plated the products of these 4 transformations strains in liquor and on solid environment with 2 antibiotics: ampicillin and chloramphenicol (Amp->LacZ; CM->AmilCP).
+
===='''1 - Ligation concentration control by Nanodrop'''====
-
<br>
+
Sheng
-
=='''Lab work'''==
+
-
<br>
+
Nanodrop :
-
==='''detail'''===
+
* BBa_K1155000 : 411.7ng/µL
-
<br>
+
===='''2 - Tranformation of BBa_K1155000 in DH5α'''====
-
*'''A.aero/anaerobic regulation system'''<br>
+
-
**''1.BioBrick promotor fnr(repressor) in plasmid PSB1C3''[[File:PS_NP.jpg|right|300px]]
+
-
:<u>Ligation product quantification</u>
+
-
:We used the nano drop to measure the DNA in 260nm and we found its concentration is 411.7ng/µl.
+
Abdou, Zhou
-
<br>
+
-
*'''A.aero/anaerobic regulation system
+
-
**''2.BioBrick RBS+LacZ+terminator in plasmid PSB1C3
+
-
**'' BioBrick RBS+amilCP+terminator in plasmid PSB1C3
+
-
<br>
+
Protocol : [[Team:Paris_Saclay/Protocols/Transformation|Bacterial transformation]]
-
:<u>Transformation</u>
+
-
:We performed 4 transformations: LacZ with plasmid PSB1A2( RBST 3K plate 2 2012 suspended in 10µl H2O) , AmilCP(with plasmid PSB1C3), the products of ligation (fnr+plasmid PSB1C3) and a control.
+
We also made liquid culture with chloramphenicol or ampicilin at 37°C of our tranformations.
 +
===='''Objective : obtaining BBa_K1155003, BBa_K1155007'''====
-
{| border="1" align="center"
+
===='''1 - Tranformation of BBa_I732017, BBa_K592009 in DH5α'''====
-
|-
+
-
|name
+
-
|LacZ in PSB1A2
+
-
|AmilCP in PSB1C3
+
-
|Mixture of ligation
+
-
|control
+
-
|-
+
-
|Vol.
+
-
|2µl
+
-
|2µL
+
-
|10µl
+
-
|10µl
+
-
|}
+
-
:And we added in each sample 50µl competent cells.
+
Abdou, Zhou
-
:After thermal shock, those sample were mixed with 1ml LB medium, incubated at 37 °C during 1h30.
+
-
<br>
+
-
<u>Cloning</u>
+
Protocol : [[Team:Paris_Saclay/Protocols/Transformation|Bacterial transformation]]
-
<br>
+
-
+
-
:We prepared Petri dishes by 2 different mediums:<br>
+
-
:*500ml LB+0.5ml chloramphenicol for AmilCP, mixture and control
+
We also made liquid culture with chloramphenicol or ampicilin at 37°C of our tranformations.
-
:*250ml LB+0.25ml ampicillin for LacZ
+
-
:Then the products of transformations were seed on the medium:<br>
 
-
:*1.Seed directly 100µL of each one on their medium
+
=='''Human practices'''==
-
:*2.Seed indirectly after concentration of products of transformation(centrifuge->remove supernatant->resuspend)
+
 
 +
We made the second meeting about open source : [[Team:Paris_Saclay/opensourcereflexion|Open Source reflexion]]
-
:The incubation lasted on night.
 
-
<br>
 
{| border="1" align="center"
{| border="1" align="center"
|[[Team:Paris Saclay/Notebook/July/1|<big>Previous day</big>]]
|[[Team:Paris Saclay/Notebook/July/1|<big>Previous day</big>]]
Line 76: Line 49:
|[[Team:Paris Saclay/Notebook/July/3|<big>Next day</big>]]
|[[Team:Paris Saclay/Notebook/July/3|<big>Next day</big>]]
|}
|}
-
 
{{Team:Paris_Saclay/incl_fin}}
{{Team:Paris_Saclay/incl_fin}}

Latest revision as of 10:06, 30 September 2013

Navigation Home Team Project Labwork Notebook Human practices

Contents

Notebook : July 2

Lab work

A - Aerobic/Anaerobic regulation system

PS NP.jpg

Objective : obtaining BBa_K1155000

1 - Ligation concentration control by Nanodrop

Sheng

Nanodrop :

  • BBa_K1155000 : 411.7ng/µL

2 - Tranformation of BBa_K1155000 in DH5α

Abdou, Zhou

Protocol : Bacterial transformation

We also made liquid culture with chloramphenicol or ampicilin at 37°C of our tranformations.

Objective : obtaining BBa_K1155003, BBa_K1155007

1 - Tranformation of BBa_I732017, BBa_K592009 in DH5α

Abdou, Zhou

Protocol : Bacterial transformation

We also made liquid culture with chloramphenicol or ampicilin at 37°C of our tranformations.


Human practices

We made the second meeting about open source : Open Source reflexion


Previous day Back to calendar Next day

Retrieved from "http://2013.igem.org/Team:Paris_Saclay/Notebook/July/2"