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Edinburgh/3 July 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
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.
