Revision as of 19:00, 9 September 2008

Transformation of OmpR*

The cloning of EnvZ* didn't worked. So we decided to try one more time, starting from the PCR product.

Digestion of EnvZ* and lasR-pSB1A2

Excision of the band corresponding to pSB1A2

Purification

EnvZ*: directly by the Qiagen kit
pSB1A2: excision of the band of interest (2 kb) and then purification using the Qiagen kit

We did again D110 and D125.

@@ Line 2: / Line 2: @@
 ='''Transformation of OmpR*'''=
-='''Construction of pFlhB - mRFP Tripart (LVA+)'''=
-Aim : Construction of ''' "pFlhB-RBS-mRFP-dbl ter" (pFlhB-I732078)''' [[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
-<br>We can only do the construction with mRFP Tripart (LVA+) because the stable strain with the Biobricks I732011 (mRFP Tripart LVA-) don't to growth.
-==Digestion==
-==='''Measurement of the concentration of D187 purified'''===
-[[Team:Paris/Notebook/Protocols#Concentration_of_the_Miniprep | Protocol (it's same that for Miniprep)]]
-{|border="1" style="text-align: center"
-|'''Digestion Name'''
-|'''Concentration (µg/mL)'''
-|'''Ratio 260/280'''
-|-
-|D187
-|16
-|1.31
-|}
-==Ligation==
-[[Team:Paris/Notebook/Protocols#Ligation |Protocol]]
-{|border="1" style="text-align: center"
-|'''Ligation Name'''
-|'''Vector Name'''
-|'''Volume Vector (µL)'''
-|'''Insert'''
-|'''Volume Insert (µL)'''
-|-
-|L168
-|D187
-|
-|D186
-|
-|-
-|Control L168
-|D166
-|
-| -
-|}
 =Cloning of EnvZ*=