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Edinburgh/23 July 2008
From 2008.igem.org
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* M43 (BABEL2+''glgC''-mut1,2) sequence results received. First and second mutation sites successfully removed. (AH) | * M43 (BABEL2+''glgC''-mut1,2) sequence results received. First and second mutation sites successfully removed. (AH) | ||
* Overnight culture of ''C. fimi'' did not yield sufficient cells. Made two more cultures in nutrient broth; all cultures incubated at 30°C with shaking. (AM) | * Overnight culture of ''C. fimi'' did not yield sufficient cells. Made two more cultures in nutrient broth; all cultures incubated at 30°C with shaking. (AM) | ||
| - | * PCR of ''crtY'' from ''P. ananatis'' cells ('''P27''') and | + | * PCR of ''crtY'' from ''P. ananatis'' cells ('''P27''') and fusion PCR of rbs+''crtB'' and rbs+''crtI'' from ligations L17 and L18 respectively ('''P28, P29'''). PCR products were run on '''Gel 22''' - P28 and P29 successful, P27 unsuccessful. (AM, Yan, OG) |
* Re-did positive control PCR of ''rrnB'' from ''C. fimi'' ('''P30''') ~ denaturing at 95°C for 20s, annealing at 55°C for 1 minute, extension at 70°C for 30s. The first two settings were in error: ''denaturing'' should have been set to 1 minute and annealing to 10 sec as usual. HOWEVER, annealing for 1 minute appears to have worked, so it might be worth trying this for PCRing the cellulases. (AM) | * Re-did positive control PCR of ''rrnB'' from ''C. fimi'' ('''P30''') ~ denaturing at 95°C for 20s, annealing at 55°C for 1 minute, extension at 70°C for 30s. The first two settings were in error: ''denaturing'' should have been set to 1 minute and annealing to 10 sec as usual. HOWEVER, annealing for 1 minute appears to have worked, so it might be worth trying this for PCRing the cellulases. (AM) | ||
* P30 (above) and pZntA were run on '''Gel 23'''. Both successful. (AM)<br /> | * P30 (above) and pZntA were run on '''Gel 23'''. Both successful. (AM)<br /> | ||
Revision as of 11:57, 26 August 2008
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Week 6
Wednesday 23 July 08
- M43 (BABEL2+glgC-mut1,2) sequence results received. First and second mutation sites successfully removed. (AH)
- Overnight culture of C. fimi did not yield sufficient cells. Made two more cultures in nutrient broth; all cultures incubated at 30°C with shaking. (AM)
- PCR of crtY from P. ananatis cells (P27) and fusion PCR of rbs+crtB and rbs+crtI from ligations L17 and L18 respectively (P28, P29). PCR products were run on Gel 22 - P28 and P29 successful, P27 unsuccessful. (AM, Yan, OG)
- Re-did positive control PCR of rrnB from C. fimi (P30) ~ denaturing at 95°C for 20s, annealing at 55°C for 1 minute, extension at 70°C for 30s. The first two settings were in error: denaturing should have been set to 1 minute and annealing to 10 sec as usual. HOWEVER, annealing for 1 minute appears to have worked, so it might be worth trying this for PCRing the cellulases. (AM)
- P30 (above) and pZntA were run on Gel 23. Both successful. (AM)
