Team:Paris Saclay/Notebook/August/21

From 2014.igem.org

(Difference between revisions)
(→D - Lemon Scent=)
m (→Electro elution of pPS1 digested by Pac1)
 
(16 intermediate revisions not shown)
Line 2: Line 2:
=Thursday 21st August=
=Thursday 21st August=
==Lab work==
==Lab work==
-
===D - Lemon Scent===
+
===Construction of the fusion protein (color)===
-
====Transformation of DH5 alpha with CAD and chromoprotein====
+
====Transformation of DH5α with chromoprotein (in pGMETeasy)====
''by Sean''
''by Sean''
 +
Petri dishes were IPTG Xgal Amp
 +
[https://2014.igem.org/Team:Paris_Saclay/Protocols/Transformation_of_competent_e.coli_by_CaCl2 protocol]
 +
 +
===Lemon Scent===
 +
====Transformation of DH5α with CAD====
 +
''by Sean''
 +
 +
[https://2014.igem.org/Team:Paris_Saclay/Protocols/Transformation_of_competent_e.coli_by_CaCl2 protocol]
 +
 +
==== Electro elution of pPS1 digested by Pac1 ====
 +
 +
[[File:Paris_Saclay_140821.jpg|left]]
 +
 +
''by Huang vu and laetitia''
 +
 +
We extracted the DNA from the agarose band saves yesterday
 +
 +
[https://2014.igem.org/Team:Paris_Saclay/Protocols/Electro_extraction_of_DNA_from_agarose_gel protocol of electro elution]
 +
 +
''Gel electrophoresis''
 +
 +
# ladder SM0403 5µl
 +
# BBa_K762100 10µl
 +
# BBa_K517003 10µl
 +
# p cola 10µl
 +
# pPSI 10µl
 +
 +
====Extraction of pPSI====
 +
'' By Mélanie''
 +
 +
From preculture of bacteria with pPSI we extract the plasmid
 +
 +
[https://2014.igem.org/Team:Paris_Saclay/Protocols/Extraction_of_the_Genomic_DNA_from_Bacteria_without_NucleoSpin%C2%AE_Tissue protocol]
 +
 +
====Extraction of pGMETeasy with LS GS and PS====
 +
 +
''By Mélanie''
 +
 +
Following the cloning made by Laeticia in pGMETeasy, i extract the plasmid ([https://2014.igem.org/Team:Paris_Saclay/Protocols/Extraction_of_the_Genomic_DNA_from_Bacteria_by_using_NucleoSpin%C2%AE_Tissue protocol])
 +
 +
and I do some stock of each stain
 +
 +
 +
==== Ligation of CAD in pMCS5 ====
 +
''by Eugene''
 +
 +
We have made an electrophoresis of purified CAD and pMCS5.
 +
 +
 +
'''Dephosphorylation of pMCS5'''
 +
 +
{| class="wikitable centre" width="50%"
 +
|+
 +
|-
 +
! scope=col | component
 +
! scope=col | volume
 +
|-
 +
|H<sub>2</sub>O
 +
|3 μl
 +
|-
 +
| pMCS5p
 +
| 5 μl
 +
|-
 +
| 10x buffer
 +
| 10 μl
 +
|-
 +
| Fast AP
 +
| 1 μl
 +
|}
 +
 +
30 min incubation in 37°c
 +
 +
15min of inactivation 65°c
 +
 +
'''Ligation'''
 +
 +
{| class="wikitable centre" width="50%"
 +
|+
 +
|-
 +
! scope=col | component
 +
! scope=col | volume
 +
|-
 +
|CADp
 +
|10 μl
 +
|-
 +
| pMCS5p
 +
| 2.5 μl
 +
|-
 +
| 10x buffer
 +
| 1.5 μl
 +
|-
 +
| Ligase
 +
| 1 μl
 +
|}
 +
 +
night at 16°c
 +
 +
 +
[https://2014.igem.org/Team:Paris_Saclay/Protocols/Electro_extraction_of_DNA_from_agarose_gel protocol]
 +
 +
==Photo of the Day==
 +
[[File:Paris Saclay 21_august.jpg|600px|center]]
 +
 +
'''Members present:'''
 +
* Instructors and advisors: Alice, Solenne and Sylvie.
 +
* Students: Eugène, Hoang Vu, Laëtitia, Mélanie, Romain, Sean and Terry.
{{Team:Paris_Saclay/notebook_footer}}
{{Team:Paris_Saclay/notebook_footer}}

Latest revision as of 15:07, 14 October 2014

Contents

Thursday 21st August

Lab work

Construction of the fusion protein (color)

Transformation of DH5α with chromoprotein (in pGMETeasy)

by Sean

Petri dishes were IPTG Xgal Amp protocol

Lemon Scent

Transformation of DH5α with CAD

by Sean

protocol

Electro elution of pPS1 digested by Pac1

Paris Saclay 140821.jpg

by Huang vu and laetitia

We extracted the DNA from the agarose band saves yesterday

protocol of electro elution

Gel electrophoresis

  1. ladder SM0403 5µl
  2. BBa_K762100 10µl
  3. BBa_K517003 10µl
  4. p cola 10µl
  5. pPSI 10µl

Extraction of pPSI

By Mélanie

From preculture of bacteria with pPSI we extract the plasmid

protocol

Extraction of pGMETeasy with LS GS and PS

By Mélanie

Following the cloning made by Laeticia in pGMETeasy, i extract the plasmid (protocol)

and I do some stock of each stain


Ligation of CAD in pMCS5

by Eugene

We have made an electrophoresis of purified CAD and pMCS5.


Dephosphorylation of pMCS5

component volume
H2O 3 μl
pMCS5p 5 μl
10x buffer 10 μl
Fast AP 1 μl

30 min incubation in 37°c

15min of inactivation 65°c

Ligation

component volume
CADp 10 μl
pMCS5p 2.5 μl
10x buffer 1.5 μl
Ligase 1 μl

night at 16°c


protocol

Photo of the Day

600px

Members present:

  • Instructors and advisors: Alice, Solenne and Sylvie.
  • Students: Eugène, Hoang Vu, Laëtitia, Mélanie, Romain, Sean and Terry.

Back to the calendar

Retrieved from "http://2014.igem.org/Team:Paris_Saclay/Notebook/August/21"