Team:Hawaii/Notebook/2008-06-13
From 2008.igem.org
(Difference between revisions)
(New page: {{Team:Hawaii/Header}} = Things we did today = == Wetlab work == ===Prepped Cells for Glycerol Stock/Seed Stock=== :<strong> Krystle</strong> :* Summary of task and what was done. Link t...) |
(→Quote of the Day) |
||
(10 intermediate revisions not shown) | |||
Line 5: | Line 5: | ||
===Prepped Cells for Glycerol Stock/Seed Stock=== | ===Prepped Cells for Glycerol Stock/Seed Stock=== | ||
:<strong> Krystle</strong> | :<strong> Krystle</strong> | ||
+ | :* Inoculated a single colony in each 10 mL LB + antibiotic<br> | ||
+ | {| border="1" | ||
+ | |+ '''Title''' | ||
+ | !Antibiotic | ||
+ | !Cell type | ||
+ | !Plasmid | ||
+ | |- | ||
+ | | rowspan="4" align="center"|Amp<sub>100</sub>||align="center"| StrataClone supercompetent cells || align="center"|pUC18 | ||
+ | |- | ||
+ | | align="center"|DH5α|| align="center"|GFP | ||
+ | |- | ||
+ | | align="center"|DH5α|| align="center"|tet+GFP | ||
+ | |- | ||
+ | | align="center"|DH5α|| align="center"|RBS (BBa_B0034) | ||
+ | |- | ||
+ | | align="center"|Kan<sub>50</sub>||align="center"|HB101|| align="center"|RK1 | ||
+ | |- | ||
+ | | align="center"|Sm<sub>2.5</sub> + Sp <sub>2.5</sub>|| align="center"|DH5α|| align="center"|pRL1383a | ||
+ | |} | ||
+ | |||
- | |||
- | |||
===Prepped Cells for Competency=== | ===Prepped Cells for Competency=== | ||
:<strong>Grace</strong> | :<strong>Grace</strong> | ||
- | :* | + | :* 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=== |
:<strong> Grace </strong> | :<strong> Grace </strong> | ||
- | :* | + | :* 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=== |
:<strong> Grace</strong> | :<strong> Grace</strong> | ||
- | + | :* (2) 300 mL batches of LB + 1.5% Agar. Stored in 4C. | |
- | :* (2) 300 mL batches of LB + 1.5% Agar | + | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
= Discussion = | = Discussion = | ||
:*We need to make more BG-ll 50x stock solution and TES 50x stock solution | :*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 = | = Quote of the Day = | ||
- | + | [[Image:DSC00746.JPG |thumb|left|375px| ''E. coli'' homes. Enough said.]][[Image:DSC00747.JPG|thumb|right|225px|Krystle & Grace confusing Norman, again. Two halves of the same person?]] | |
{{Team:Hawaii/Footer}} | {{Team:Hawaii/Footer}} | ||
__NOTOC__ | __NOTOC__ |
Latest revision as of 23:11, 14 June 2008
Projects | Events | Resources | ||
---|---|---|---|---|
Sponsors | Experiments | Milestones | Protocols | |
Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Prepped Cells for Glycerol Stock/Seed Stock
- Krystle
- Inoculated a single colony in each 10 mL LB + antibiotic
- Inoculated a single colony in each 10 mL LB + antibiotic
Antibiotic | Cell type | Plasmid |
---|---|---|
Amp100 | StrataClone supercompetent cells | pUC18 |
DH5α | GFP | |
DH5α | tet+GFP | |
DH5α | RBS (BBa_B0034) | |
Kan50 | HB101 | RK1 |
Sm2.5 + Sp 2.5 | DH5α | pRL1383a |
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.
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 ]