Team:Paris Saclay/Notebook/August/21

From 2014.igem.org

(Difference between revisions)
(Ligation of CAD in pMCS5)
(Transformation of DH5α with chromoprotein)
Line 3: Line 3:
==Lab work==
==Lab work==
===B - Construction of the fusion protein (color)===
===B - Construction of the fusion protein (color)===
-
====Transformation of DH5α with 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]
[https://2014.igem.org/Team:Paris_Saclay/Protocols/Transformation_of_competent_e.coli_by_CaCl2 protocol]

Revision as of 09:18, 26 August 2014

Contents

Thursday 21st August

Lab work

B - Construction of the fusion protein (color)

Transformation of DH5α with chromoprotein (in pGMETeasy)

by Sean

Petri dishes were IPTG Xgal Amp protocol

D - Lemon Scent

Transformation of DH5α with CAD

by Sean

protocol

Electro elution of pPS1 digested by Pac1

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

Paris Saclay 140821.jpg

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

Back to the calendar