IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Output/2009/07/08/Zhangs

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Transformation for the ligation product: T7p+2M/1H/11N/2G/2I/5N/1J RBS+CI+1-23L, 7 samples in total.

Double digestion for 1-2K+CI+1-23L insert again (yesterday failed)

1.5μL XbaI
1.5μL PstI
2μL 10×M buffer
10μL 1-2K③ plasmid
5μL ddH2O
20μL Total

Electrophoresis and Gel Extraction for 1-2K+CI+1-23L and CI+1-23L insert (for 1-5J vector)

2K+CI+1-23L 0.081g 243μL
CI+1-23L for 5J 0.100g 300μL

Ligate the 1-2K+CI+1-23L insert and T7 promoter vector
System

3μL 1-2K+CI+1-23L insert
1μL T7p vector
1μL 10× Ligase buffer
1μL Ligase
4μL ddH2O
10μL Total

Ligate the CI+1-23L insert and 1-5J RBS vector
System

3μL CI+1-23L insert
1μL 1-5J vector
1μL 10× Ligase buffer
1μL Ligase
4μL ddH2O
10μL Total