August/20 August 2009

1.digestion and ligation

digestion with restriction enzyme

K204007, (K204003)

K204008

↓ 37°C, 2hr

↓ gel electrophoresis UP  : 1-2M, 1-14F , 1-2M , 2-8O DOWN : No.3-A,B,C,D, No.4-A,B,C,D , No.6-A,B,C,D , Vector gel cut ↓ purification by [QIAquick Nucleotide Removal Kit] ↓ ligation ligation DNA       44 10* buffer 5 ligation  1 total     50uL ↓ 16°C overnight

2. rapid check

③(RBS+cinR),K204003 ③-C,D : OK

④(RBS+LasR),K204004 ④-A,B,C : OK

⑥(RBS+cinI),K204006 ⑥-A,B,C : OK

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