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Notebook : July 3

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining BBa_K1155000

**1 - Colony PCR of BBa_K1155000 to check good insertion of Pndh* in pSB1C3**

Abdou, Sheng, Zhou

Tranformation of 07/02/13 works. We will do a PCR colony.

Colonies count for BBa_K1155000 :

Standard concentration :
- BBa_K1155000 : 0 colony

High concentration :
- BBa_K1155000 : 2 colonies

Primers and PCR :

VF2, VR, Pfnr_up, Pfnr_down are four oligos that we used for plasmid amplification. We used tree combinaisons VF/VR, VF/Pfnr_Down, Pfnr_Up/VR. If the promoter Pndh* insert pSB1C3 plasmid successfully, tree fragments with specific size will be amplified.

Protocol

We resuspend each colony with 20µL of H2O.

Used quantities :

DNA : 2µL
Mix : (it was divided in tubes for 4 different colonies for each oligo combinaison with 23µL of mix in each tube)
- VF or Pfnr_Up : 6µL
- VR or Pfnr_Down : 6µL
- dNTP 10mM : 6µL
- Buffer Dream Taq : 30µL
- Dream Taq : 6µL
- H2O : 246µL

2 - Culture of BBa_K1155000

Sheng

We made new cultures by streaking 2 colonies of 07/02/13 culture of BBa_K1155000. We also did liquid culture of each one. We incubate culture at 37°C.

Objective : obtaining BBa_K1155003, BBa_K1155007

1 - Culture of BBa_I732017, BBa_K592009

Sheng

Tranformation of 07/02/13 works. We will do new cultures.

We made new cultures by streaking 2 colonies of 07/02/13 culture of BBa_I732017, BBa_K592009. We also did liquid culture of each one. We incubate culture at 37°C.

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@@ Line 1: / Line 1: @@
 {{Team:Paris_Saclay/incl_debut_generique}}
 ='''Notebook : July 3'''=
 =='''Lab work'''==
-constructing
+==='''A - Aerobic/Anaerobic regulation system'''===
+===='''Objective : obtaining BBa_K1155000'''====
+===='''1 - Colony PCR of BBa_K1155000 to check good insertion of Pndh* in pSB1C3'''====
+Abdou, Sheng, Zhou
+{|
+| style="border:1px solid black;padding:5px;background-color:#DE;" |
+Tranformation of 07/02/13 works. We will do a PCR colony.
+|}
+Colonies count for BBa_K1155000 :
+* Standard concentration :
+** BBa_K1155000 : 0 colony
+* High concentration :
+** BBa_K1155000 : 2 colonies
+[[File:Ps0307jour.jpg|300px]]
 <br>
+<center>[[File:PSprimer07.jpg|right|250px]]</center>
+<br>
+'''[https://2013.igem.org/Team:Paris_Saclay/Primers Primers] and PCR :'''
+<p>'''VF2, VR, Pfnr_up, Pfnr_down are four oligos that we used for plasmid amplification. We used tree combinaisons VF/VR, VF/Pfnr_Down, Pfnr_Up/VR.'''
+'''If the promoter Pndh* insert pSB1C3 plasmid successfully, tree fragments with specific size will be amplified.''' </p>
+'''
+Protocol'''
+We resuspend each colony with 20µL of H2O.
+Used quantities :
+* DNA : 2µL
+* Mix : (it was divided in tubes for 4 different colonies for each oligo combinaison with 23µL of mix in each tube)
+** VF or Pfnr_Up : 6µL
+** VR or Pfnr_Down : 6µL
+** dNTP 10mM : 6µL
+** Buffer Dream Taq : 30µL
+** Dream Taq : 6µL
+** H2O : 246µL
+[[File:PSPCR0307.jpg|400px]]
+===='''2 - Culture of BBa_K1155000 '''====
+Sheng
+We made new cultures by streaking 2 colonies of 07/02/13 culture of BBa_K1155000. We also did liquid culture of each one.
+We incubate culture at 37°C.
+===='''Objective : obtaining BBa_K1155003, BBa_K1155007'''====
+===='''1 - Culture of BBa_I732017, BBa_K592009 '''====
+Sheng
+{|
+| style="border:1px solid black;padding:5px;background-color:#DE;" |
+Tranformation of 07/02/13 works. We will do new cultures.
+|}
+We made new cultures by streaking 2 colonies of 07/02/13 culture of BBa_I732017, BBa_K592009. We also did liquid culture of each one.
+We incubate culture at 37°C.
 {| border="1" align="center"
 |[[Team:Paris Saclay/Notebook/July/2|<big>Previous day</big>]]
-|[[Team:Paris_Saclay/Notebook|<big>Back to calender</big>]]
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Team:Paris Saclay/Notebook/July/3

From 2013.igem.org