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 |
HB101 |
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
[http://manoa.hawaii.edu/ 
][http://manoa.hawaii.edu/ovcrge/ 
][http://www.ctahr.hawaii.edu 
]
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