IGEM:Peking/2007/Count-Conjugation-Notebook/2007-7-19

Miniprep

• For I7100-2;
• Use Transgen Kit;
• Final volume: 10mL

Digestion Test: I7100-2

• I7100-2 .
• Digesting I7100 with EcoRI and EcoRI/XbaI.
• Digestion system contains (totally 20uL):

1. single digestion using EcoRI: 2uL 10*H, 0.5uL EcoRI, 6uL plasmid, 12uL ddH2O.
2. double digestion using EcoRI/XbaI:2uL 10*M, 1uL EcoRI, 1uL XbaI, 6uL plasmid, 10uL ddH2O.

Electrophoresis result:

• from Left to Right:

1. I7100-2 @ EcoRI
2. I7100-2 @ EcoRI/XbaI
3. I7100-2 plasmid
4. Marker

• notes:**************

Digestion Test: E0040 & R0040

• test if the new Dye helps to improve the resolution.
• Digesting E0040 with EcoRI and EcoRI/PstI.
• Digesting R0040 with EcoRI.

1. single digestion using EcoRI: 2uL 10*H, 0.5uL EcoRI, 6uL plasmid, 11.5uL ddH2O.
2. double digestion using EcoRI/XbaI:2uL 10*H, 0.5uL EcoRI, 0.5uL XbaI, 6uL plasmid, 11uL ddH2O.

Electrophoresis result:

• from Left to Right:

1. E0040 @ EcoRI
2. E0040 @ EcoRI/PstI
3. E0040-1 plasmid
4. R0040 @ EcoRI
5. R0040-1 plasmid
6. I7100 @ EcoRI
7. I7100 @ EcoRI/XbaI
8. I7100-2 plasmid
9. Marker

Electrophoresis - control

• from Left to Right:

1. E0040 @ EcoRI
2. E0040 @ EcoRI/PstI
3. R0040 @ EcoRI
4. E0040-1 plasmid
5. R0040-1 plasmid
6. Marker

conclusion

• @@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@

mimiprep

• E0040-3, E0040-4, E0040-5
• R0040-2, R0040-3, R0040-4