Team:Chiba/Notebook 3
From 2010.igem.org
Characterization of ''lux'' inverter
2010-10-8
transformation Plasmid:BBa_R0061 BBa_E0240 strain:XL10-G
2010-10-9
Mini-prep (Sigma Aldrich) BBa_R0061 BBa_E0240 digestion -------------------- BBa_R0061 20 µL 10×Buffer 3 µL SpeI 1 µL PstI 1 µL NFW 5 µL -------------------- 30 µL -------------------- BBa_E0240 20 µL 10×Buffer 3 µL XbaI 1 µL PstI 1 µL NFW 5 µL -------------------- 30 µL ↓ gel extraction
2010-10-11
Ligation ----------------------- BBa_R0061 (70 ng) 1 µL BBa_E0240 (100 ng) 2 µL ATP (10 mM) 1 µL 10×Buffer 1 µL T4 DNA Ligase 1 µL NFW 4 µL ----------------------- 10 µL ↓ transformation strain:XL10-G
2010-10-12
Ligation ----------------------- BBa_R0061(100 ng) 1 µL BBa_E0240 (100 ng) 3 µL ATP (10 mM) 1 µL T4 DNA Ligase 1 µL dH<sub>2</sub>O 4 µL ----------------------- 10 µL ↓ transformation strain:XL10-Gold
2010-10-21
Double transformation Strain:DH10B Plasmid:Plux inv-GFP & Plac-LuxR
2010-10-22
liquid culture LB (Carb-Kan) 1mL 37℃ 12 h 200 rpm
2010-10-26