IGEM:Cambridge/2008/Notebook/Bacillus/2008/08/15: Difference between revisions
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- Plate out on Amp 100 (Neat) | - Plate out on Amp 100 (Neat) | ||
===Biobrick R0010, E0040 and I13522=== | |||
Revision as of 10:38, 19 September 2008
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Transformation ECE188, 189, 190 into E.coli- Results : only our control plate grew!! - Maybe we did not add enough DNA, or there is another problem. We will e mail the guy from Bacillus center before trying again, because we don't have enough DNA. Plasmid miniprep ECE 151 (x2), ECE 153, ECE 166
Xylose Induction with ECE 153- 4 Tubes - No fluorescence! We have to think about the transformation of integration vectors into Bacillus.
Glycerol Stock Transformation- Spin down cells (competent) from 5/8 and 13/8 stocks - Remove liquid, resuspend in Medium B - Incubate for 60 mins at 37°C - Add ECE 166 into tubes - Incubate for 30 mins at 37°C - Plate out on CM 5 plates - 4 plates
Transforming ECE 188, 189, 190 (3rd try)- 2 tubes of chemically competent top 10 - Spin down, remove liquid - Resuspend cells in 100μL of CaCl2 50mM - 4 tubes with 50μL of cells
- Ice for 30 mins - 42°C for 2 mins - Ice for 2 mins - Incubate at 37°C for 2 hours - Plate out on Amp 100 (Neat) Biobrick R0010, E0040 and I13522 |
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