IGEM:Cambridge/2008/Notebook/Bacillus/2008/08/07

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Transformation of B.S. IA771

New transformation with the same protocol than 2 days ago. We will try to add less liquid on plates (to avoid growing colonies in liquid)

Tube Plate 1 Plate 2 Plate 3 Plate 4
ECE166 Cm5 + 200μL of cells Cm5 + 100μL of cells Cm5 + 50μL of cells Cm10 + 100μL of cells
ECE153 Spc50 + 200μL of cells Spc50 + 100μL of cells Spc50 + 50μL of cells
ECE166 Cm5 + 100μL of cells Cm10 + 100μL of cells
ECE153 Spc50 + 100μL of cells
no DNA Cm5 + 100μL of cells Cm10 + 100μL of cells Spc50 + 100μL of cells
ECE166 (glycerol stock) Cm5 + 100μL of cells Cm10 + 100μL of cells

Check Vectors

We want to make a final check for ECE147, 149, 150, 153 and 162.

- Double digest + gel

  • Results
  • Lane3 : HyperladderI
  • Lane4 : ECE147
  • Lane5 : ECE149
  • Lane6 : ECE150
  • Lane7 : ECE153
  • Lane8 : ECE162
  • Lane9 : HyperladderI
Photom.gif

- ECE 147, ECE150, ECE153 :ok!

- ECE 149 : 3 bands! not the right vector

- ECE162 : ony one band! It has been uncut... maybe the good vector (according to precedent gel), but not sure

Test ECE112 transformation

- for the three different strain (1A1, IA771, IA751), make 12 spots

- take one colony with a loop, and put it on a Cm5 plate and on a Cm5 + Spc100 plate (do that for the 12 spots)

- incubate

New stock

- Put ECE171 in 10mL of LB

- Incubate