Paris/October 20

[[Paris/October 19|yesterday]] -- [[Paris/October 21|tomorrow]] <br>
== Digestion Reaction ==

== PCR reaction ==

{{Paris_PCR_0| Title = PrefixBB-frt1-CmR-frt2-SufixBB
|Denaturation= 98°C 2' first / 20" then
|Annealing= /
|Elongation= 72°C 1'40"
|Cycles= 30
|Final elongation= 72°C 10'
|Buffer=  (5x) 10µL
|MgCl2= 10µM 0µL (no extra)
|dNTP= 10µM 1µL
|n_oligoF= O54
|v_oligoF= 10µM 4µL
|n_oligoR= O55
|v_oligoR= 10µM 4µL
|water= qsp 50µL
|pol= Phusion 0,5µL
|DNA= pY-2P-IntC 5µL
|Size= 1000bp
|Success=YES
|Image=
|Band=
|}}

{{Paris_PCR_0| Title = PrefixBB-frt1-CmR-frt2-SufixBB
|Denaturation= 98°C 2' first / 20" then
|Annealing= /
|Elongation= 72°C 1'40"
|Cycles= 30
|Final elongation= 72°C 10'
|Buffer=  (5x) 10µL  ! GC one !
|MgCl2= 10µM 0µL (no extra)
|dNTP= 10µM 1µL
|n_oligoF= O54
|v_oligoF= 10µM 4µL
|n_oligoR= O55
|v_oligoR= 10µM 4µL
|water= qsp 50µL
|pol= Phusion 0,5µL
|DNA= pY-2P-IntC 5µL
|Size= 1000bp
|Success=NO
|Image=
|Band=
|}}

{{Paris_PCR_0| Title = PrefixBB-frt1-CmR-frt2-SufixBB
|Denaturation= 98°C 2' first / 20" then
|Annealing= /
|Elongation= 72°C 1'40"
|Cycles= 30
|Final elongation= 72°C 10'
|Buffer=  (5x) 10µL ! GC one !
|MgCl2= 10µM 0µL (no extra)
|DMSO= 2,5µL (or 5%)
|dNTP= 10µM 1µL
|n_oligoF= O54
|v_oligoF= 10µM 4µL
|n_oligoR= O55
|v_oligoR= 10µM 4µL
|water= qsp 50µL
|pol= Phusion 0,5µL
|DNA= pY-2P-IntC 5µL
|Size= 1000bp
|Success=NO
|Image=
|Band=
|}}

{{Paris_PCR_0| Title = PrefixBB-frt1-CmR-frt2-T1T2-ufixBB
|Denaturation= 98°C 2' first / 20" then
|Annealing= /
|Elongation= 72°C 1'40"
|Cycles= 30
|Final elongation= 72°C 10'
|Buffer=  (5x) 10µL
|MgCl2= 10µM 0µL (no extra)
|dNTP= 10µM 1µL
|n_oligoF= O54
|v_oligoF= 10µM 4µL
|n_oligoR= O56
|v_oligoR= 10µM 4µL
|water= qsp 50µL
|pol= Phusion 0,5µL
|DNA= pY-2P-IntC 5µL
|Size= 1000bp
|Success=YES
|Image=
|Band=
|}}

{{Paris_PCR_0| Title = PrefixBB-frt1-CmR-frt2-T1T2-SufixBB
|Denaturation= 98°C 2' first / 20" then
|Annealing= /
|Elongation= 72°C 1'40"
|Cycles= 30
|Final elongation= 72°C 10'
|Buffer=  (5x) 10µL  ! GC one !
|MgCl2= 10µM 0µL (no extra)
|dNTP= 10µM 1µL
|n_oligoF= O54
|v_oligoF= 10µM 4µL
|n_oligoR= O56
|v_oligoR= 10µM 4µL
|water= qsp 50µL
|pol= Phusion 0,5µL
|DNA= pY-2P-IntC 5µL
|Size= 1000bp
|Success=NO
|Image=
|Band=
|}}

{{Paris_PCR_0| Title = PrefixBB-frt1-CmR-frt2-T1T2-SufixBB
|Denaturation= 98°C 2' first / 20" then
|Annealing= /
|Elongation= 72°C 1'40"
|Cycles= 30
|Final elongation= 72°C 10'
|Buffer=  (5x) 10µL ! GC one !
|MgCl2= 10µM 0µL (no extra)
|DMSO= 2,5µL (or 5%)
|dNTP= 10µM 1µL
|n_oligoF= O54
|v_oligoF= 10µM 4µL
|n_oligoR= O56
|v_oligoR= 10µM 4µL
|water= qsp 50µL
|pol= Phusion 0,5µL
|DNA= pY-2P-IntC 5µL
|Size= 1000bp
|Success=YES
|Image=
|Band=
|}}

== Purification of Digestion and PCR products ==

== Ligation Reactions ==

{|
|- style="background: #ccccff;"
! colspan="5" style="background: #ccccff;" | Ligations
|- style="background: #ccccff; text-align: center;"
|width=5%| Number
|width=20%| Insert
|width=5%| Insert Volume (µL)
|width=25%| Vector
|width=5%| Vector Volume (µL)
|width=35%| Comments
|width=5%| Number of colonies
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |C22
|
|
|D47
|3
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |C23
|
|
|D72
|3
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |C24
|
|
|D72 (from Oct. 14)
|3
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |C25
|
|
|D50
|3
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |C26
|
|
|D41
|3
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |C27
|
|
|D22
|5
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |C28
|
|
|D81
|11
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |C29
|
|
|D89
|3
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |C30
|
|
|D90
|3
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |L68
|D56 - araC/pBad>>RBS-Cre BBinsert
|10
|D72 - pSB2K3 BBvector
|3
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |L68 bis
|D56 (14/10/07) - araC/pBad>>RBS-Cre BBinsert
|10
|D72 (14/10/07) - pSB2K3 BBvector
|3
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |L69
|D92
|10
|D50
|3
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |L70
|D92
|10
|D41
|3
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |L71
|D93
|15
|D50
|3
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |L72
|D93
|15
|D41
|3
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |L73
|D84
|10
|D22
|5
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |L74
|D80
|3
|D81
|15
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |L74 bis
|D80
|4
|D81
|12
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |L75
|D83
|4
|D89
|3
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |L76
|D83
|10
|D90
|3
|
|0
|- style="background: #cccccc;"  
| style="background: #ccffcc;" |L77
|D94.6
|3
|D95
|10
|
|0
|}

== Culture ==

Launching of 80mL culture of the strain countaining the plasmid pY-2P-IntC, last minipreps revealed the weak nomber of plasmid recover with a classic miniprep on 5mL culture.