"

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
Sample:Plux inv-GFP and Plac-LuxR

LB (Carb-Kan,AHL 0 nM and 1000 nM ) 1mL
37℃ 12 h 200 rpm

2010-10-26

Spin-dawn and observed the pellet