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