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Edinburgh/8 July 2008
From 2008.igem.org
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Week 4
Tuesday 8 July 08
- New primers for crt BioBricks arrived. Performed PCR with KOD using primer pairs crtBf2 (new) + crtBr2 (old)(P8), crtIf2 + crtIr2 (both new) (P9), and crtYf2 (new) + crtYr2 (old)(P10). The template for P8 and P10 was a suspension of Pantoea ananatis cells from plate 17; for P9, Douglas's maxiprep DNA from 19 Feb (with the PstI sites mutated out) was used. In all cases annealing was at 55°C and extension for 30s. Gel 9 showed that P8 and P9 had worked, with pure products of the expected size seen, but no product was seen for P10.
- glgC mutagenesis: Transformed competent cells with ligation products L6 and L7; plated cells on Plates 22 and 23 (L6: BABEL2+glgC-mut1) and Plates 24 and 25 (L7: BABEL2+glgC-mut2).
- Ran gel 9 to check size of X1 (J33201) and X2 (pSB1A2+dxs) HindIII restriction digest. The digest looked fine.
