IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Output/2009/06/26/Zhangs

Miniprep 12 samples:
CI
Terminators: 1-23L, 1-4H
RBS: 1-2I, 1-2G, 1.2M, 1-5J, 1-5N, 1-1J, 1-1H, 1-2K, 1-11N
The 12 plasmid samples stored in -20°C

Enzymatic digestion
Samples: CI, 1-23L, 1-4H
System:
For CI insert:

5μL	plasmid
5μL	10×H buffer
1μL	EcoRI
1μL	SpeI
38μL	ddH2O
20μL	Total

For terminator vectors 1-23L & 1-4H:

5μL	plasmid
5μL	10×H buffer
1μL	EcoRI
1μL	XbaI
38μL	ddH2O
20μL	Total

Add CIAP into 1-4H & 1-23L vector digestion product.
Electrophoresis the digested product.
Samples loading order: CI1, CI2, M, CI plasmid (Control)

Result reveals that the double-digestion failed. The double-digested fragments can’t be identified out in electrophoresis causing the concentration of plasmids in this enzymatic digestion system is too low.

Put 3 samples (CI, I-4H, 1-23L) into the shaker