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