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Team:Hawaii/Notebook/2008-10- 2
From 2008.igem.org
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Verification of transformants
- Grace
File:100208colonyPCR.png
EtBr stained 2% agarose gel ran at 95V for 1 hour. Two microliters of PCR product were loaded into each well.
| Construct | Colonies |
|---|---|
| pSB1A3, SAP'd | 0 |
| pSB1A3 to self | 40 |
| plac + rbs+GFP | 42 |
| rbs + slr1+GFP | 89 |
| rbs+GFP + tt | 227 |
| slr1+GFPf + tt | 178 |
| nir + rbs+GFP | 112 |
| nir+rbs + slr1+GFP | 215 |
| nir + rbs+GFP+tt #1 | 300+ |
| nir + rbs+GFP+tt #2 | 103 |
| plac + rbs+GFP+tt #1 | 63 |
| plac + rbs+GFP+tt #2 | 11 |
- Colony PCR of transformants\
- Most lanes blank; lanes with bands are mostly faint. Didn't load enough PCR product? Will reload gel tomorrow with remainder of PCR product.
- Restreaked:
- rbs+GFP+tt #9, 10, 16, 17
- slr1+GFPf+tt #13
- nir+rbs+slr1+GFPf #1, 2, 15, 17
Plasmid Prep
- Margaret
- oriV 3, 4, & 6
Restriction Digest
- Margaret
- oriV3 E,S
- oriV4 E,S
- oriV6 E,S
- oriV11 E,S
- plac/rbs/rep 8 X,P
- pSB1A3 E,P
- oriT E,S, and another sample of oriT with X,P
Culture
- Margaret
- ap1, and ap2. Because I need to digest them with E,S so that I can use as a front insert
PCR Verification
- Margaret
- 10 more colonies of plac/rbs/rep
- check if SAP and SAP buffer is contaminated
Sequencing
- Margaret
- oriV 3, 4, 6
- plac/rbs/rep 8 with 11 primers to cover whole sequence
Discussion
Quote of the Day
History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson
[http://manoa.hawaii.edu/ 
][http://manoa.hawaii.edu/ovcrge/ 
][http://www.ctahr.hawaii.edu 
]
