Team:Hawaii/Notebook/2008-10- 2
From 2008.igem.org
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(→Verification of transformants) |
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::* slr1+GFPf+tt #13 | ::* slr1+GFPf+tt #13 | ||
::* nir+rbs+slr1+GFPf #1, 2, 15, 17 | ::* nir+rbs+slr1+GFPf #1, 2, 15, 17 | ||
+ | |||
+ | ===Plasmid Prep=== | ||
+ | :<strong> Margaret</strong> | ||
+ | :*oriV 3, 4, & 6 | ||
+ | |||
+ | ===Restriction Digest=== | ||
+ | :<strong> Margaret </strong> | ||
+ | :*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=== | ||
+ | :<strong> Margaret </strong> | ||
+ | :*ap1, and ap2. Because I need to digest them with E,S so that I can use as a front insert | ||
+ | |||
+ | ===PCR Verification=== | ||
+ | :<strong> Margaret </strong> | ||
+ | |||
+ | :*10 more colonies of plac/rbs/rep | ||
+ | :*check if SAP and SAP buffer is contaminated | ||
+ | |||
+ | |||
+ | ===Sequencing=== | ||
+ | :<strong> Margaret </strong> | ||
+ | |||
+ | :*oriV 3, 4, 6 | ||
+ | :*plac/rbs/rep 8 with 11 primers to cover whole sequence | ||
+ | |||
= Discussion = | = Discussion = |
Latest revision as of 04:33, 3 October 2008
Projects | Events | Resources | ||
---|---|---|---|---|
Sponsors | Experiments | Milestones | Protocols | |
Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Verification of transformants
- Grace
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 ]