Revision as of 19:07, 16 September 2008

Purification of yesterday digestion

D112 before gel excision

Digestion n°	DNA substrat	Digestion by
D112	MP106: S03879 (rbs-TetR) in pSB1A2	EcoRI & SpeI

1% agarose gel

Qiaquick Gel Extraction Kit

Sample	DNA concentration	A260/A280
D112	8 µg/mL	2,31

Defroze of the cells on ice
Add 5 µL of DNA in 100 µL of competent cells (ligation products or pUC19 for positive control)
30 min on ice
Heat schock 45sec at 42°C
2 min on ice
Add 900 µL of SOC
Incubate 1h at 37°C
Centrifugate 5 min at 6000 rpm
Remove 800 µL
Plate the 200 µL left on LB + amplicilline
incubate O/N at 37°C

@@ Line 51: / Line 51: @@
-=Construction of pFlhB-mRFP-LVA-tripart & pFliL-ECFP-LVA-tripart=
+=Constructions=
 [[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
-*L183 = pFlhB - mRFP LVA+
+==List of the constructions used to transformate==
-*L184 = pFliL - ECFP LVA+
+{|border="1" style="text-align: center"
-*L185 = pFliL - ECFP LVA-
+|'''Ligation n°'''
+|'''Insert'''
+|'''Vector'''
+|'''Construction'''
+|-
+|L173
+|D112
+|D187
+|rbs-tetR-mRFP-LVA+
+|-
+|L183
+|D134
+|D187
+| pFlhB - mRFP LVA+
+|-
+|L184
+|D149
+|D198
+| pFliL - ECFP LVA+
+|-
+|L185
+|D149
+|D199
+| pFliL - ECFP LVA-
+|}