IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Input/2009/07/11/Shuke Wu

Gel electrophoresis (again)
Products of double digest of L1, L2, t2, t3,
Marker: 100bp 250bp 500bp 750bp 1kb 2kb 3kb 5kb
Loading buffer and DNA dye: 6×
Voltage and time: 60V 5min; 120V 15min
Lane1: L1: insert 1.1kb;
Lane2: L2: insert 1.1kb;
Lane3: marker;
Lane5: t2: insert 700bp;
Lane6: t3: insert 700bp;

DNA ligation
System 10uL: Insert 6uL, vector 2uL, buffer 1uL, T4 DNA ligase 1uL
16℃ 4 hour
Insert: L1 L2 t2 t3;
Vector: terminator digested by EcoR1 and Xba1 (provided by Haoqian Zhang & Guosheng Zhang)

Transformation (by Min Lin)
Products of ligation, competent cells 50uL each,
Smear to LB plate with Amp