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 - 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