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

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Contents

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

Image:2007-7-19_I7100-2_EcoRI+XbaI.jpg

  • 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

Image:2007-7-19_E0040-1_EcoRI_and_EcoRI+PstI_R0040-1_EcoRI_I7100-2_EcoRI_and_EcoRI+XbaI.jpg‎

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

Image:2007-7-19_E0040-1_EcoRI_and_EcoRI+PstI_R0040-1_EcoRI(control).jpg‎

conclusion

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

mimiprep

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