IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/07/25

Antoine

Minipreps

• B0015 272.5 ng/μL
• J23110 991.9 ng/μL

Restriction digest

• B0015 cut by X+P: 25 μL miniprep (22/07) + 1 μL each enzyme + 0.3 μL BSA + 3 μL Buffer 3
• P1003 cut by S+P: 6 μL miniprep (22/07) + 1 μL each enzyme + 0.1 μL BSA + 1 μL Buffer 2 + 0.9 H20
• J23110 cut by S+P: 16 μL miniprep (22/07) + 1 μL each enzyme + 0.2 μL BSA + 2 μL Buffer 2
• C0040 cut by X+P: 9 μL miniprep (22/07) + 1 μL each enzyme + 0.15 μL BSA + 1.5 μL Buffer 3 + 2.35 H20
• C0012 cut by X+P: 19 μL miniprep (22/07) + 1 μL each enzyme + 0.25 μL BSA + 2.5 μL Buffer 3 + 1.25 H2O

Inactivation 20 min 65°C and storage at -20°C for J23110 digest product.

Electrophoresis Gel 2%

Raphaël
Glycerol 15% 2 glycerols per culture, one stored at -20°C and the other at -80°C

• TOP10 pSB1A2 weak RBS -TetA(C) - Amp
• TOP10 pSB1A2 med.RBS -TetA(C) - Amp
• TOP10 pSB1A2 weak RBS -LuxR - Amp
• TOP10 pSB1A2 med.RBS -LuxR - Amp
• TOP10 pSB1A2 stand.RBS -LuxR - Amp

Minipreps : 4 minipreps of a 10ml culture per biobrick, eluted with 30μL buffer and collected into 1 tube for a total volume of around 120μL:

• weak RBS -TetA(C) - Amp
• med.RBS -TetA(C) - Amp
• weak RBS -LuxR - Amp
• med.RBS -LuxR - Amp
• stand.RBS -LuxR - Amp

Nanodrop to determine the amount of DNA in the miniprep samples: