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Edinburgh/1 August 2008
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<li><a href="https://2008.igem.org/Team:Edinburgh" title="Home"><span>Home</span></a></li> | <li><a href="https://2008.igem.org/Team:Edinburgh" title="Home"><span>Home</span></a></li> | ||
<li><a href="https://2008.igem.org/Team:Edinburgh/Project" title="Project" rel="dropmenu1_a"><span>The Project</span></a></li> | <li><a href="https://2008.igem.org/Team:Edinburgh/Project" title="Project" rel="dropmenu1_a"><span>The Project</span></a></li> | ||
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<li><a href="https://2008.igem.org/Team:Edinburgh/Modeling" title="Modelling" rel="dropmenu2_a"><span>Modelling</span></a></li> | <li><a href="https://2008.igem.org/Team:Edinburgh/Modeling" title="Modelling" rel="dropmenu2_a"><span>Modelling</span></a></li> | ||
| - | <li><a href="https://2008.igem.org/Team:Edinburgh/Notebook" title="Notebook"><span>Notebook</span></a></li> | + | <li><a href="https://2008.igem.org/Team:Edinburgh/Notebook" title="Notebook"><span>Notebook</span></a></li> |
| + | <li><a href="https://2008.igem.org/Team:Edinburgh/Results" title="Results" rel="dropmenu1_a"><span>Results</span></a></li> | ||
| + | <li><a href="http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2008&group=Edinburgh" title="Parts" rel="dropmenu1_a"><span>BioBrick Parts</span></a></li> | ||
| + | <li><a href="https://2008.igem.org/Team:Edinburgh/Team" title="Team" ><span>The Team</span></a></li> | ||
</ul> | </ul> | ||
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| + | |align="left" width="150pt"|{{#calendar: title=Edinburgh |year=2008 | month=06}} | ||
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| + | |align="left" width="150pt"|{{#calendar: title=Edinburgh |year=2008 | month=07}} | ||
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| + | |align="left" width="150pt"|{{#calendar: title=Edinburgh |year=2008 | month=08}} | ||
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| + | :::: '''[[Edinburgh/31_July_2008|< Previous Entry]]''' | ||
=== Week 7 === | === Week 7 === | ||
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* Primers for pCstA ordered. (CF) | * Primers for pCstA ordered. (CF) | ||
| - | * White colonies from Thursday's transformations were subplated as follows | + | * White colonies from Thursday's transformations were subplated as follows: |
** Plate 76 (L26: pSB1A2+rbs+''appY'') to '''Plate 83''' | ** Plate 76 (L26: pSB1A2+rbs+''appY'') to '''Plate 83''' | ||
** Plate 77 (L27: pSB1A2+rbs+''crtB'') to '''Plate 84''' | ** Plate 77 (L27: pSB1A2+rbs+''crtB'') to '''Plate 84''' | ||
** Plate 80 (L28: pSB1A2+''cenA'') to '''Plate 85''' | ** Plate 80 (L28: pSB1A2+''cenA'') to '''Plate 85''' | ||
** Plates 81/82 (L29: pSB1A2+''cex'') to '''Plate 86'''. (AM/HX/OG) | ** Plates 81/82 (L29: pSB1A2+''cex'') to '''Plate 86'''. (AM/HX/OG) | ||
| - | * ''First digestion'': M68 | + | * ''First digestion'': M68-M71 (pSB1A2+''glgC''-mut1,2) with EcoRI/PstI, M72-M75 (pSB1A2+rbs+''dxs'') with EcoRI, M76-M79 (BABEL2+''crtE'') with EcoRI/PstI. Run on '''Gel 33'''. Results: M68, 69, 71: vector band around 2 kb, no ''glgC'' band; M72, 73, 74: single band around 4 kb, expected length for vector+gene; M76-79: ''crtE'' band <1 kb, vector band ~2 kb. It is not clear into which vector ''crtE'' (M76-79) is inserted, so sequencing will be necessary. (AM) |
| - | * ''Second digestion'': M68 and M69 with a) EcoRI, b) EcoRI/PstI; M72 with a) EcoRI b) SacI/SpeI. Run on '''Gel 34'''. Results: Single and double digests of M68 and M69 yielded similar results, hence pSB1A2+''glgC''-mut1,2 ligation or transformation failed; digests of M72 yielded expected bands, hence pSB1A2+rbs+''dxs'' successful. (AM) | + | * ''Second digestion'': M68 and M69 with a) EcoRI, b) EcoRI/PstI; M72 with a) EcoRI b) SacI/SpeI. Run on '''Gel 34'''. Results: Single and double digests of M68 and M69 yielded similar results, hence pSB1A2+''glgC''-mut1,2 ligation or transformation failed; digests of M72 yielded expected bands, hence pSB1A2+rbs+''dxs'' probably successful. (AM) |
| - | * P36 (''glgC''-mut1,2 excision from BABEL2) and L25 (P36 + Edinbrick1) were run again on '''Gel 35'''. Results: P36 viewed at lower illumination yields one band of the desired length (1.5kb); L25 yields two bands around 1.5kb, a thick band at 3kb and several fainter bands including one at 2kb (length of pSB1A2). (AM) | + | * P36 (''glgC''-mut1,2 excision from BABEL2) and L25 (P36 + Edinbrick1) were run again on '''Gel 35'''. Results: P36 viewed at lower illumination yields one band of the desired length (1.5kb); L25 yields two bands around 1.5kb, a thick band at 3kb and several fainter bands including one at 2kb (length of pSB1A2). (AM)<br /> |
| + | <br /> | ||
| + | |||
| + | :::: '''[[Edinburgh/1_August_2008|Next Entry >]]''' | ||
Revision as of 16:14, 27 August 2008
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Week 7
Friday 1 August 08
- Primers for pCstA ordered. (CF)
- White colonies from Thursday's transformations were subplated as follows:
- Plate 76 (L26: pSB1A2+rbs+appY) to Plate 83
- Plate 77 (L27: pSB1A2+rbs+crtB) to Plate 84
- Plate 80 (L28: pSB1A2+cenA) to Plate 85
- Plates 81/82 (L29: pSB1A2+cex) to Plate 86. (AM/HX/OG)
- First digestion: M68-M71 (pSB1A2+glgC-mut1,2) with EcoRI/PstI, M72-M75 (pSB1A2+rbs+dxs) with EcoRI, M76-M79 (BABEL2+crtE) with EcoRI/PstI. Run on Gel 33. Results: M68, 69, 71: vector band around 2 kb, no glgC band; M72, 73, 74: single band around 4 kb, expected length for vector+gene; M76-79: crtE band <1 kb, vector band ~2 kb. It is not clear into which vector crtE (M76-79) is inserted, so sequencing will be necessary. (AM)
- Second digestion: M68 and M69 with a) EcoRI, b) EcoRI/PstI; M72 with a) EcoRI b) SacI/SpeI. Run on Gel 34. Results: Single and double digests of M68 and M69 yielded similar results, hence pSB1A2+glgC-mut1,2 ligation or transformation failed; digests of M72 yielded expected bands, hence pSB1A2+rbs+dxs probably successful. (AM)
- P36 (glgC-mut1,2 excision from BABEL2) and L25 (P36 + Edinbrick1) were run again on Gel 35. Results: P36 viewed at lower illumination yields one band of the desired length (1.5kb); L25 yields two bands around 1.5kb, a thick band at 3kb and several fainter bands including one at 2kb (length of pSB1A2). (AM)
- Next Entry >
