Team:Paris/August 1

Strong rbs - lasR activator
D129 (BV) - D114 (BI)

Strong promoter - ECFP
D123 (BV) - D130 (BI)

Strong promoter - gfp Tripart
D123 (BV) - D131 (BI)

Strongest promoter - ECFP
D103 (BV) - D131 (BI)

Strong promoter - gfp Tripart
D103 (BV) - D130 (BI)

Medium promoter - gfp Tripart
D124 (BV) - D131 (BI)

Weak promoter - ECFP
D104 (BV) - D130 (BI)

Weak promoter - gfp
D104 (BV) - D131 (BI)

pLas - ECFP
D105 (BV) - D130 (BI)

pLas - gfp Tripart
D105 (BV) - D131 (BI)

Double Terminator - yfp
D125 (FV) - D117 (FI)

Double Terminator - rfp
D125 (FV) - D121 (FI)

Double Terminator - lasR activator + LVA
D125 (FV) - D113 (FI)

Strong promoter - gfp tripart
D106 (BV) - D130 (BI)

Strongest RBS - rfp
D102 (BV) - D122 (BI)

Strongest RBS - yfp
D102 (BV) - D118 (BI)

Transformations

Protocol

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

List of transformation