Edinburgh/3 July 2008
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* Clones M2, M10 and M11 were submitted for sequencing using primers pSB1A2f1 and pSB1A2r1 (for M2) and pTG262f1 and pTG262r1 (for M10 and M11). We should get the results on Monday. | * Clones M2, M10 and M11 were submitted for sequencing using primers pSB1A2f1 and pSB1A2r1 (for M2) and pTG262f1 and pTG262r1 (for M10 and M11). We should get the results on Monday. | ||
* Ordered primers to make carotenoid biosynthesis BioBricks:<br /> | * Ordered primers to make carotenoid biosynthesis BioBricks:<br /> | ||
- | ::: '''primer crtIf2''': gat gaattc gcggccgc t tctag atg aaa cca act acg gta att g<br /> | + | :::: '''primer crtIf2''': gat gaattc gcggccgc t tctag atg aaa cca act acg gta att g<br /> |
- | ::: 22 matches, 8 GC = 32 C, 14 AT = 28 C, total 60°C, total length = 45<br /> | + | :::: 22 matches, 8 GC = 32 C, 14 AT = 28 C, total 60°C, total length = 45<br /> |
- | ::: 83.2°C, moderate, no<br /> | + | :::: 83.2°C, moderate, no<br /> |
- | ::: '''primer crtIr2''': gct actagt a tta tt a tat cag atc ctc cag cat c<br /> | + | :::: '''primer crtIr2''': gct actagt a tta tt a tat cag atc ctc cag cat c<br /> |
- | ::: 20 matches, 9 GC = 36, 11 AT = 22, total 58°C, length = 35<br /> | + | :::: 20 matches, 9 GC = 36, 11 AT = 22, total 58°C, length = 35<br /> |
- | ::: 66.1°C, weak, no<br /> | + | :::: 66.1°C, weak, no<br /> |
- | ::: '''primer crtBf2''': gat gaattc gcggccgc t tctag atg aat aat ccg tcg tta ctc<br /> | + | :::: '''primer crtBf2''': gat gaattc gcggccgc t tctag atg aat aat ccg tcg tta ctc<br /> |
- | ::: 21 matches, 8 GC = 32, 13 AT = 26, total 58°C. length = 44<br /> | + | :::: 21 matches, 8 GC = 32, 13 AT = 26, total 58°C. length = 44<br /> |
- | ::: 82.9°C, moderate, no<br /> | + | :::: 82.9°C, moderate, no<br /> |
- | ::: '''primer crtYf2''': gat gaattc gcggccgc t tctag atg caa ccg cat tat gat ctg<br /> | + | :::: '''primer crtYf2''': gat gaattc gcggccgc t tctag atg caa ccg cat tat gat ctg<br /> |
- | ::: 21 matches, 9 GC = 36,12 AT = 24, total 60°C, length = 44<br /> | + | :::: 21 matches, 9 GC = 36,12 AT = 24, total 60°C, length = 44<br /> |
- | ::: 86.4°C, very strong, no<br /> | + | :::: 86.4°C, very strong, no<br /> |
: Note that we already have a compliant ''crtE'' BioBrick (I hope), and the reverse primers crtBr2 and crtYr2 should be fine (see iGEM2007 lab book, page 90). | : Note that we already have a compliant ''crtE'' BioBrick (I hope), and the reverse primers crtBr2 and crtYr2 should be fine (see iGEM2007 lab book, page 90). | ||
* Retransformed JM109 with the remainder of the ''appY'' ligation (I re-ligated it, and also heat-treated it before the transformation, since Tom Knight reports that this can increase transformation efficiency). Plated this to '''Plates 15 and 16'''.<br /> | * Retransformed JM109 with the remainder of the ''appY'' ligation (I re-ligated it, and also heat-treated it before the transformation, since Tom Knight reports that this can increase transformation efficiency). Plated this to '''Plates 15 and 16'''.<br /> |
Revision as of 10:13, 21 August 2008
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Week 3
Thursday 3 July 08
- Clones M2, M10 and M11 were submitted for sequencing using primers pSB1A2f1 and pSB1A2r1 (for M2) and pTG262f1 and pTG262r1 (for M10 and M11). We should get the results on Monday.
- Ordered primers to make carotenoid biosynthesis BioBricks:
- primer crtIf2: gat gaattc gcggccgc t tctag atg aaa cca act acg gta att g
- 22 matches, 8 GC = 32 C, 14 AT = 28 C, total 60°C, total length = 45
- 83.2°C, moderate, no
- primer crtIr2: gct actagt a tta tt a tat cag atc ctc cag cat c
- 20 matches, 9 GC = 36, 11 AT = 22, total 58°C, length = 35
- 66.1°C, weak, no
- primer crtBf2: gat gaattc gcggccgc t tctag atg aat aat ccg tcg tta ctc
- 21 matches, 8 GC = 32, 13 AT = 26, total 58°C. length = 44
- 82.9°C, moderate, no
- primer crtYf2: gat gaattc gcggccgc t tctag atg caa ccg cat tat gat ctg
- 21 matches, 9 GC = 36,12 AT = 24, total 60°C, length = 44
- 86.4°C, very strong, no
- primer crtIf2: gat gaattc gcggccgc t tctag atg aaa cca act acg gta att g
- Note that we already have a compliant crtE BioBrick (I hope), and the reverse primers crtBr2 and crtYr2 should be fine (see iGEM2007 lab book, page 90).
- Retransformed JM109 with the remainder of the appY ligation (I re-ligated it, and also heat-treated it before the transformation, since Tom Knight reports that this can increase transformation efficiency). Plated this to Plates 15 and 16.