IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Output/2009/06/24/Shan Shen

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10:50 Miniprep the plasmids containing T7 promoter, standard parts 1-12M and 1-12O.
1-12M: medium rbs + GFP coding gene + terminator
1-12O: strong rbs + GFP coding gene + terminator 12:30 Digest the plasmids of T7 promoter, 1-12M and 1-12O
Digestion System for plasmids of T7 promoter:50μL

Plasmids 30μL
Spe1 1μL
Pst1 1μL
Buffer 1*H 5μL.

Digestion System for plasmids of 1-12M and 1-12O: 50μL

Plasmids 30μL
Xba1 1μL
Pst1 1μL
Buffer 1*M 5μL

12:50 Start to digest
14:20 Add CIAP into the system of vector digestion.

Buffer 5μL
Enzyme 1μL

15:20 Electrophoresis to recycle the vectors and inserts of digestion.
The order of the samples: marker, plasimid control of T7 promoter, T7 promoter, 1-12M, 1-12O.
PKU 20090624 Shan Shen 1.JPG
Recycle the fragments (vector) of T7 promoter and the inserts of 1-12M, 1-12O.
The fragment of T7 promoter is weird, for the linear plasmids ran even faster than the round ones. Decide to do some controls and tests