Team:Paris/July 30

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Revision as of 09:17, 5 August 2008

Results of transformations

Ligation name	Description	Antibio	Nb colonies observed	Comments
L100	rbs TetR - ECFP D110 (BV) - D130 (BI)	Amp	0	After another night nothing was osberved -> To do again
L101	rbs TetR - GFP tripart D110 (BV) - D131 (BI)	Amp	1	After another night nothing was observed -> To do again
L102	Strong rbs - YFP D129 (BV) - D118 (BI)	Amp	+/- 100
L103	Strong rbs - GFP D129 (BV) - D122 (BI)	Amp	a lot
L104	Strong rbs - lasR activator D129 (BV) - D114 (BI)	Amp	a lot
L105	Strong promoter - ECFP D123 (BV) - D130 (BI)	Amp	a lot
L106	Strong promoter - gfp Tripart D123 (BV) - D131 (BI)	Amp	a lot
L107	Strongest promoter - ECFP D103 (BV) - D131 (BI)	Amp	a lot
L108 n°2 (the right one)	Strong promoter - gfp Tripart D103 (BV) - D130 (BI)	Amp	+/- 100
L109 n°1	Strong promoter - ecfp D124 (BV) - D130 (BI)		+/- 30	To do again
L109 n°2	Strong promoter - ecfp D124 (BV) - D130 (BI)		a lot	To do again
L110	Medium promoter - gfp Tripart D124 (BV) - D131 (BI)	Amp	a lot
L111	Weak promoter - ECFP D104 (BV) - D130 (BI)	Amp	a lot
L112	Weak promoter - gfp D104 (BV) - D131 (BI)	Amp	a lot
L113	AracpBAD - ecfp D126 (BV) - D130 (BI)	Kan	1	After another night nothing was observed -> To do again
L114	AracpBAD - gfp tripart D126 (BV) - D131 (BI)	Kan	2	After another night nothing was observed -> To do again
L115	pLas - ECFP D105 (BV) - D130 (BI)	Amp	a lot
L116	pLas - gfp Tripart D105 (BV) - D131 (BI)	Amp	a lot
L117	yfp - Double Terminator D117 (FI) - D125 (FV)	Amp	+/- 30
L118	rfp - Double Terminator D121 (FI) - D125 (FV)	Amp	+/- 30
L119	lasR activator + LVA - Double Terminator D113 (FI) - D125 (FV)	Amp	+/- 30
L120	tetR repressible - ECFP D106 (BV) - D130 (BI)	Amp	a lot
L121	Strong promoter - gfp tripart D106 (BV) - D130 (BI)	Amp	a lot
L122	RBS-lasI - ecfp D107 (BV) - D130 (BI)	Amp	0		After another night nothing was observed -> To do again
L123	RBS lasI - ecfp D107 (BV) - D131 (BI)	Amp	1		After another night nothing was observed -> To do again
L124	Strongest RBS - rfp D102 (BV) - D122 (BI)	Amp	a lot	To do again
L125	Strongest RBS - yfp D102 (BV) - D118 (BI)		a lot ++		To do again
L126	Strongest RBS (1)- LacR activator (+LVA) D102 (BV) - D118 (BI)		no dish found!!!		To do again
L127	gfp (1)- Double terminator D119 (FI) - D125 (FV)		no dish found!!!		To do again
Controls
C1	D110		0		-
C2	D129		5		-
C3	D123		+/- 100		-
C4	D103		+/- 20		-
C5	D124		+/- 100		-
C6	D104		+/- 100		-
C7	D126		0		-
C8	D105		+/- 20		-
C9	D125		0		-
C10	D106		0		-
C11	D107		0		-
C12	D102		+/- 10		-
Positive control	puc19		155 (transformation efficiency:1.5*10^7/ug)		-

Analysis of yesterday DNA digestion

The digested DNA of yesterday was analysed one more time by electrophoresis on a 0.8% agarose gel (about 30 minutes at 100 W). The ladder used was the 1 kb DNA ladder (New England Biolabs). 5 µL of each sample with 1 µL of loading dye were loaded.

D101	B0034	1	EcoRI	XbaI	FV	2076 bp	2000 bp	6	10
Name	BioBrick	Tube N°	Enz 1	Enz 2	Obs	Exp Size	Mea Size	Conc (ng/µl)	Band
D102	B0034	1	SpeI	PstI	BV	2077 bp	2000 bp	8	11
D105	R0079	1	SpeI	PstI	BV	2222 bp	2000 bp	6	13
D106	R0040	1	SpeI	PstI	BV	2119 bp	2000 bp	6	12
D108	S03154	1	XbaI	PstI	BI	707 bp		0 (failed)	14
D111	S03879	1	XbaI	PstI	BI	725 bp	600 bp	2	15
D115	C0179	2	EcoRI	SpeI	FI	746 bp		0 (failed)	4 & 5
D116	C0179	2	XbaI	PstI	BI	745 bp	700 bp	2	2 & 3
D117	E0030	1	EcoRI	SpeI	FI	746 bp	600 bp	10	16
D128	B0030	2	EcoRI	XbaI	FV	2079 bp	2000 bp	8	6 & 7
D129	B0030	2	SpeI	PstI	BV	2080 bp	2000 bp	8	8 & 9

Results : Each of the samples was succesfully digested and purified except for the sample D108. It seems that the QIAprep columms (from the QIAGEN Minipreps kit) can be used instead of the QIAquick columms (for DNA Gel Extraction).

@@ Line 1: / Line 1: @@
 {{Paris/Calendar_Links|July 29|July 31}}
+== Results of transformations ==
+{| border="1"
+|align="center"|'''Ligation name'''
+|align="center"|'''Description'''
+|align="center"|'''Antibio'''
+|align="center"|'''Nb colonies observed'''
+|align="center"|'''Comments'''
+|-
+|align="center"|L100
+|align="center"|
+rbs TetR - ECFP
+<br>D110 (BV) -  D130 (BI)
+|align="center"|Amp
+|align="center"|0
+|align="center"|After another night nothing was osberved -> To do again
+|-
+|align="center"|L101
+|align="center"|
+rbs TetR - GFP tripart
+<br>D110 (BV) -  D131 (BI)
+|align="center"|Amp
+|align="center"|1
+|align="center"|After another night nothing was observed -> To do again
+|-
+|align="center"|L102
+|align="center"|
+Strong rbs - YFP
+<br>D129 (BV) -  D118 (BI)
+|align="center"|Amp
+|align="center"|+/- 100
+|align="center"|
+|-
+|align="center"|L103
+|align="center"|
+Strong rbs - GFP
+<br>D129 (BV) -  D122 (BI)
+|align="center"|Amp
+|align="center"|a lot
+|align="center"|
+|-
+|align="center"|L104
+|align="center"|
+Strong rbs - lasR activator
+<br>D129 (BV) -  D114 (BI)
+|align="center"|Amp
+|align="center"|a lot
+|align="center"|
+|-
+|align="center"|L105
+|align="center"|
+Strong promoter - ECFP
+<br>D123 (BV) - D130 (BI)
+|align="center"|Amp
+|align="center"|a lot
+|align="center"|
+|-
+|align="center"|L106
+|align="center"|
+Strong promoter - gfp Tripart
+<br>D123 (BV) - D131 (BI)
+|align="center"|Amp
+|align="center"|a lot
+|align="center"|
+|-
+|align="center"|L107
+|align="center"|
+Strongest promoter - ECFP
+<br>D103 (BV) - D131 (BI)
+|align="center"|Amp
+|align="center"|a lot
+|align="center"|
+|-
+|align="center"|L108 n°2 (the right one)
+|align="center"|
+Strong promoter -  gfp Tripart
+<br>D103 (BV) - D130 (BI)
+|align="center"|Amp
+|align="center"|+/- 100
+|align="center"|
+|-
+|align="center"|L109 n°1
+|align="center"|
+Strong promoter -  ecfp
+<br>D124 (BV) - D130 (BI)
+|align="center"|
+|align="center"|+/- 30
+|align="center"|To do again
+|-
+|align="center"|L109 n°2
+|align="center"|
+Strong promoter -  ecfp
+<br>D124 (BV) - D130 (BI)
+|align="center"|
+|align="center"|a lot
+|align="center"|To do again
+|-
+|align="center"|L110
+|align="center"|
+Medium promoter - gfp Tripart
+<br>D124 (BV) - D131 (BI)
+|align="center"|Amp
+|align="center"|a lot
+|align="center"|
+|-
+|align="center"|L111
+|align="center"|
+Weak promoter - ECFP
+<br>D104 (BV) - D130 (BI)
+|align="center"|Amp
+|align="center"|a lot
+|align="center"|
+|-
+|align="center"|L112
+|align="center"|
+Weak promoter - gfp
+<br>D104 (BV) - D131 (BI)
+|align="center"|Amp
+|align="center"|a lot
+|align="center"|
+|-
+|align="center"|L113
+|align="center"|
+AracpBAD - ecfp
+<br>D126 (BV) - D130 (BI)
+|align="center"|Kan
+|align="center"|1
+|align="center"|After another night nothing was observed -> To do again
+|-
+|align="center"|L114
+|align="center"|
+AracpBAD - gfp tripart
+<br>D126 (BV) - D131 (BI)
+|align="center"|Kan
+|align="center"|2
+|align="center"|After another night nothing was observed -> To do again
+|-
+|align="center"|L115
+|align="center"|
+pLas - ECFP
+<br>D105 (BV) - D130 (BI)
+|align="center"|Amp
+|align="center"|a lot
+|align="center"|
+|-
+|align="center"|L116
+|align="center"|
+pLas - gfp Tripart
+<br>D105 (BV) - D131 (BI)
+|align="center"|Amp
+|align="center"|a lot
+|align="center"|
+|-
+|align="center"|L117
+|align="center"|
+yfp - Double Terminator
+<br>D117 (FI) - D125 (FV)
+|align="center"|Amp
+|align="center"|+/- 30
+|align="center"|
+|-
+|align="center"|L118
+|align="center"|
+rfp - Double Terminator
+<br>D121 (FI) - D125 (FV)
+|align="center"|Amp
+|align="center"|+/- 30
+|align="center"|
+|-
+|align="center"|L119
+|align="center"|
+lasR activator + LVA - Double Terminator
+<br>D113 (FI) - D125 (FV)
+|align="center"|Amp
+|align="center"|+/- 30
+|align="center"|
+|-
+|align="center"|L120
+|align="center"|
+tetR repressible - ECFP
+<br>D106 (BV) - D130 (BI)
+|align="center"|Amp
+|align="center"|a lot
+|align="center"|
+|-
+|align="center"|L121
+|align="center"|
+Strong promoter - gfp tripart
+<br>D106 (BV) - D130 (BI)
+|align="center"|Amp
+|align="center"|a lot
+|align="center"|
+|-
+|align="center"|L122
+|align="center"|
+RBS-lasI - ecfp
+<br>D107 (BV) - D130 (BI)
+|align="center"|Amp
+|align="center"|0
+|align="center"|
+|align="center"|After another night nothing was observed -> To do again
+|-
+|align="center"|L123
+|align="center"|
+RBS lasI - ecfp
+<br>D107 (BV) - D131 (BI)
+|align="center"|Amp
+|align="center"|1
+|align="center"|
+|align="center"|After another night nothing was observed -> To do again
+|-
+|align="center"|L124
+|align="center"|
+Strongest RBS - rfp
+<br>D102 (BV) - D122 (BI)
+|align="center"|Amp
+|align="center"|a lot
+|align="center"|To do again
+|-
+|align="center"|L125
+|align="center"|
+Strongest RBS - yfp
+<br>D102 (BV) - D118 (BI)
+|align="center"|
+|align="center"|a lot ++
+|align="center"|
+|align="center"|To do again
+|-
+|align="center"|L126
+|align="center"|
+Strongest RBS (1)- LacR activator (+LVA)
+<br>D102 (BV) - D118 (BI)
+|align="center"|
+|align="center"|no dish found!!!
+|align="center"|
+|align="center"|To do again
+|-
+|align="center"|L127
+|align="center"|
+gfp (1)- Double terminator
+<br>D119 (FI) - D125 (FV)
+|align="center"|
+|align="center"|no dish found!!!
+|align="center"|
+|align="center"|To do again
+|-
+|align="center"|'''Controls'''
+|align="center"|
+|align="center"|
+|align="center"|
+|align="center"|
+|align="center"|
+|-
+|align="center"|C1
+|align="center"|
+D110
+|align="center"|
+|align="center"|0
+|align="center"|
+|align="center"|-
+|-
+|align="center"|C2
+|align="center"|
+D129
+|align="center"|
+|align="center"|5
+|align="center"|
+|align="center"|-
+|-
+|align="center"|C3
+|align="center"|
+D123
+|align="center"|
+|align="center"|+/- 100
+|align="center"|
+|align="center"|-
+|-
+|align="center"|C4
+|align="center"|
+D103
+|align="center"|
+|align="center"|+/- 20
+|align="center"|
+|align="center"|-
+|-
+|align="center"|C5
+|align="center"|
+D124
+|align="center"|
+|align="center"|+/- 100
+|align="center"|
+|align="center"|-
+|-
+|align="center"|C6
+|align="center"|
+D104
+|align="center"|
+|align="center"|+/- 100
+|align="center"|
+|align="center"|-
+|-
+|align="center"|C7
+|align="center"|
+D126
+|align="center"|
+|align="center"|0
+|align="center"|
+|align="center"|-
+|-
+|align="center"|C8
+|align="center"|
+D105
+|align="center"|
+|align="center"|+/- 20
+|align="center"|
+|align="center"|-
+|-
+|align="center"|C9
+|align="center"|
+D125
+|align="center"|
+|align="center"|0
+|align="center"|
+|align="center"|-
+|-
+|align="center"|C10
+|align="center"|
+D106
+|align="center"|
+|align="center"|0
+|align="center"|
+|align="center"|-
+|-
+|align="center"|C11
+|align="center"|
+D107
+|align="center"|
+|align="center"|0
+|align="center"|
+|align="center"|-
+|-
+|align="center"|C12
+|align="center"|
+D102
+|align="center"|
+|align="center"|+/- 10
+|align="center"|
+|align="center"|-
+|-
+|align="center"|Positive control
+|align="center"|
+puc19
+|align="center"|
+|align="center"|155 (transformation efficiency:1.5*10^7/ug)
+|align="center"|
+|align="center"|-
+|}
 ==Analysis of yesterday DNA digestion==