From 2008.igem.org
Things we did today
Wetlab work
Prepped Cells for Glycerol Stock/Seed Stock
- Krystle
Inoculated a single colony in each 10 mL LB + antibiotic
<tbody>
| Antibiotic |
Cell
Type |
Plasmid |
| Amp100 |
supercompetent cells |
pUC18 |
| DH5alpha |
GFP |
| DH5alpha |
tet+GFP |
| DH5alpha |
RBS |
| Kan50 |
E.coli |
RK1 |
| Sm2.5
+ Sp 2.5 |
DH5alpha |
pRL1383a |
</tbody>
Prepped Cells for Competency
- Grace
- Checked yesterday's inoculation. OD600=1.2 (too high). Need to redo.
- Made SOB for new inoculation.
- Inoculated 250 μl SOB with 100 μl DH5α. Inoculated 250 μl SOB with 10 μl DH5α. Incubated overnight at 27C with shaking (150 rpm).
Cell density test
- Grace
- Retrieved plates from SC incubator. All plates had too many colonies (lawn). Need to redo.
- Made BG-11 plates for retest next week.
Made LB agar stock
- Grace
- (2) 300 mL batches of LB + 1.5% Agar. Stored in 4C.
Drylab Work
Discussion
- We need to make more BG-ll 50x stock solution and TES 50x stock solution
- Kelly from SC lab will be in 12pm-3pm. We should retrieve our triparental conjugation plates then.
- Grace, Krystle, and Norman will come in at 9am tomorrow to plate triparental conjugates, make competent cells, and glycerol stock purified Biobrick colonies.
Quote of the Day
History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson
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