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Team:Hawaii/Notebook/2008-10- 8
From 2008.igem.org
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Triparental conjugation (cont.)
- Grace
- PCR'd BBpRL cultures to ensure that plasmid was retained
- Plasmid was not retained (grown in SOB w/o selection = no reason to retain plasmid)
- PCR'd BBpRL colonies #1 and 18 from plate to ensure plasmid present
- Regrew RP1, BBpRL harboring cultures in 20ml SOB with selection (Kan50 and Sp100 respectively) and incubated at 37C overnight, with shaking (rpm = 250)
Verification of transformants
- Grace
| Construct | Colonies |
|---|---|
| nir + rgt#1 | 1 |
| plac + rgt#1 | 0 |
| nr + sg | 12 |
| pr + sg | 74 |
| nr + pgt | 5 |
| pr + pgt | 19 |
| pSB1A3 (control) | 1 |
| no DNA | 0 |
- Colony PCR of transformants
- Restreaked nrsg #6, prpgt #5, 7, 11
Construction of secretion device (cont.)
- Grace
- PCR'd and verified on EtBr stained 2% agarose gel:
- J33207
- nir+rbs
- nir
- rgt#1
- Overnight RE digest:
- HindIII/BamHI:
- J33207
- EcoRI/SpeI:
- nir
- nir+rbs
- XbaI/PstI:
- rgt#1
Prepared for sequencing
- Grace
- Colony PCR'd:
- pgt #21
- nrsg #1, 2, 15, 17 (from 10/2)
- sgt #13 (from 10/2)
- ExoSAP'd:
- pgt #21
- nrsg #1, 2, 15, 17 (from 10/2)
- nrsg #6 (from 10/8)
- sgt #13 (from 10/2)
- prpgt #5, 7, 11
Drylab work
iGEM title/abstract
- Grace & Margaret
- Revised abstract
- Sent in abstract, title, track selection to iGEM HQ
Discussion
Quote of the Day
History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson
