IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Input/2009/07/07/Shuke Wu

Gel electrophoresis
Products of double digest of lacI and araC,
marker: 100bp 250bp 500bp 750bp 1kb 2kb 3kb 5kb
loading buffer and DNA dye: 6×
voltage and time: 60V 5min; 120V 15min
lane1: lacI plasmid;
lane2: araC plasmid;
lane3: marker;
lane4: digested product of lacI;
lane5: digested product of araC;

DNA Gel purification
lacI and araC

PCR product purification
6 RBS

DNA ligation
System 10uL: Insert 4uL, vector 1uL, water 3uL, buffer 1uL, T4 DNA ligase 1uL
16℃ 4 hour
Insert: lacI; araC; tetR (from Min Lin);
Vertor: 6 RBS

Transformation
Products of ligation, competent cells 50uL each,
Smear to LB plate with Amp