Team:Paris/August 1

Revision as of 11:10, 5 August 2008 by Audrey (Talk | contribs)

MiniPreps

Name	Description	Biobricks
MP L102		Strong rbs - YFP D129 (BV) - D118 (BI)
MP L103		Strong rbs - GFP D129 (BV) - D122 (BI)
MP L104		Strong rbs - lasR activator D129 (BV) - D114 (BI)
MP L105		Strong promoter - ECFP D123 (BV) - D130 (BI)
MP L106		Strong promoter - gfp Tripart D123 (BV) - D131 (BI)
MP L107		Strongest promoter - ECFP D103 (BV) - D131 (BI)
MP L108		Strong promoter - gfp Tripart D103 (BV) - D130 (BI)
MP L110		Medium promoter - gfp Tripart D124 (BV) - D131 (BI)
MP L111		Weak promoter - ECFP D104 (BV) - D130 (BI)
MP L112		Weak promoter - gfp D104 (BV) - D131 (BI)
MP L115		pLas - ECFP D105 (BV) - D130 (BI)
MP L116		pLas - gfp Tripart D105 (BV) - D131 (BI)
MP L117		Double Terminator - yfp D125 (FV) - D117 (FI)
MP L118		Double Terminator - rfp D125 (FV) - D121 (FI)
MP L119		Double Terminator - lasR activator + LVA D125 (FV) - D113 (FI)
MP L121		Strong promoter - gfp tripart D106 (BV) - D130 (BI)
MP L124		Strongest RBS - rfp D102 (BV) - D122 (BI)
MP L125		Strongest RBS - yfp D102 (BV) - D118 (BI)

Strain	Ligation	Description
S123	L104	Strong rbs - lasR activator D129 (BV) - D114 (BI)
S124	L105	Strong promoter - ECFP D123 (BV) - D130 (BI)
S125	L106	Strong promoter - gfp Tripart D123 (BV) - D131 (BI)
S126	L107	Strongest promoter - ECFP D103 (BV) - D131 (BI)
S127	L108	Strong promoter - gfp Tripart D103 (BV) - D130 (BI)
S128	L110	Medium promoter - gfp Tripart D124 (BV) - D131 (BI)
S129	L111	Weak promoter - ECFP D104 (BV) - D130 (BI)
S130	L112	Weak promoter - gfp D104 (BV) - D131 (BI)
S131	L115	pLas - ECFP D105 (BV) - D130 (BI)
S132	L116	pLas - gfp Tripart D105 (BV) - D131 (BI)
S133	L117	Double Terminator - yfp D125 (FV) - D117 (FI)
S134	L118	Double Terminator - rfp D125 (FV) - D121 (FI)
S135	L119	Double Terminator - lasR activator + LVA D125 (FV) - D113 (FI)
S136	L121	Strong promoter - gfp tripart D106 (BV) - D130 (BI)
S137	L124	Strongest RBS - rfp D102 (BV) - D122 (BI)
S138	L125	Strongest RBS - yfp D102 (BV) - D118 (BI)

use of TOP10 Chemically competent cells

Defroze competent cells on ice during 5'
Add 5µl of DNA Ligation in 50µL of competent bacterias (or 1µL for the positive control puc19)
Incubate 30' on ice
Heat-shock the cells during 30" at 42°C without shaking
Put 2' on ice
Add 250µL of pre-warmed SOC medium (4°C)
Incubate 1h at 37°C under shaking (225rpm)
Spin at 5.000rpm during 30"
Remove 150µL of supernatant
Resuspent the pellet in the 150µL left
Spread on adequated plates
Incubate O/N at 37°C