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Team:Hawaii/Notebook/2008-08-17
From 2008.igem.org
(Difference between revisions)
(→Verification of transformants) |
(→Verification of transformants) |
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| Line 14: | Line 14: | ||
| align=center|BB-pRL1383a + J33207 | | align=center|BB-pRL1383a + J33207 | ||
| align=center|LB + sp<sub>100</sub> + sm<sub>50</sub> + IPTG (1mM) + X-gal (2mM) | | align=center|LB + sp<sub>100</sub> + sm<sub>50</sub> + IPTG (1mM) + X-gal (2mM) | ||
| - | |align=center| | + | |align=center|105 |
| - | |align=center| | + | |align=center|0 |
|- | |- | ||
| align=center|BB-pRL1383a + J33207 | | align=center|BB-pRL1383a + J33207 | ||
|align=center|LB + sp<sub>100</sub> + X-gal (2mM) | |align=center|LB + sp<sub>100</sub> + X-gal (2mM) | ||
| - | |align=center| | + | |align=center|44 |
| - | |align=center| | + | |align=center|0 |
|- | |- | ||
|align=center| BB-pRL1383a (negative control) | |align=center| BB-pRL1383a (negative control) | ||
|align=center|LB + sp<sub>100</sub> + sm<sub>50</sub> + IPTG (1mM) + X-gal (2mM) | |align=center|LB + sp<sub>100</sub> + sm<sub>50</sub> + IPTG (1mM) + X-gal (2mM) | ||
| - | |align=center| | + | |align=center|44 |
| - | |align=center| | + | |align=center|0 |
|- | |- | ||
| align=center|no DNA (negative control) | | align=center|no DNA (negative control) | ||
|align=center|LB + sp<sub>100</sub> + sm<sub>50</sub> + IPTG (1mM) + X-gal (2mM) | |align=center|LB + sp<sub>100</sub> + sm<sub>50</sub> + IPTG (1mM) + X-gal (2mM) | ||
| - | |align=center| | + | |align=center|0 |
| - | |align=center| | + | |align=center|0 |
|} | |} | ||
:* Colony PCR to verify insert | :* Colony PCR to verify insert | ||
| + | ::* ~325bp band is due to pRL1383a plasmid | ||
| + | ::* BB-pRL1383a + J33207 bands are smaller than (-) control bands, potential success in replacing GFP w/ lac device | ||
| + | :::* (-) bands due to contamination (no template added to PCR rxn) and GFP (religated plasmid, one RE did not cut) | ||
| + | :* Restreaked colonies to purify | ||
| + | :* Grew up colonies on TB+sp<sub>100</sub> for plasmid prep | ||
= Discussion = | = Discussion = | ||
Revision as of 00:23, 18 August 2008
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Verification of transformants
- Grace
| DNA | Plate | Total colony forming units | Blue colonies |
|---|---|---|---|
| BB-pRL1383a + J33207 | LB + sp100 + sm50 + IPTG (1mM) + X-gal (2mM) | 105 | 0 |
| BB-pRL1383a + J33207 | LB + sp100 + X-gal (2mM) | 44 | 0 |
| BB-pRL1383a (negative control) | LB + sp100 + sm50 + IPTG (1mM) + X-gal (2mM) | 44 | 0 |
| no DNA (negative control) | LB + sp100 + sm50 + IPTG (1mM) + X-gal (2mM) | 0 | 0 |
- Colony PCR to verify insert
- ~325bp band is due to pRL1383a plasmid
- BB-pRL1383a + J33207 bands are smaller than (-) control bands, potential success in replacing GFP w/ lac device
- (-) bands due to contamination (no template added to PCR rxn) and GFP (religated plasmid, one RE did not cut)
- Restreaked colonies to purify
- Grew up colonies on TB+sp100 for plasmid prep
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 
]
