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Edinburgh/30 July 2008
From 2008.igem.org
Week 7
Wednesday 30 July 08
- P48 (crtY) and P49 (Recreation of P12, rbs+dxs) created (30.07.08: CF)
- Ligated F1 (putative cenA) and F2 (putative cex) to Edinbrick 1 (L28 and L29 respectively). All were digested with EcoRI/SpeI, purified and stored overnight in the 16°C water bath. (AM)
- Repeated PCR for cenA and cex (P46, P47) from heat killed cell solution. Denaturing 95°C for 1 min, annealing 65°C for 10s, extension 70°C for 40s. Run on Gel 30. Unsuccessful, perhaps because annealing temperature was too high (temperature was decided based on stock solution label, which took the prefix/suffix into consideration). (AM)
- Gel 31: M49 and M50 (pSB1A2-rbs+crtI digested with a) XbaI, b) SpeI/XbaI, c) sac/speI), M63 (BABEL2+rbs+crtE digested with a) EcoRI, b) EcoRI/PstI), M67 (pSB1A2+rbs+crtB digested with a) EcoRI/PstI, b) sacI/speI) and P48 (crtY). (OG)
- Re-digested M55 to M60 (pSB1A2+pZntA) with a) EcoRI and b) EcoRI/PstI. (This time, the amount of DNA for digestion is increased). (Yan)
- Gel 32 results look promising for M57, but not for the others. (Yan)
- Purified P15 (rbs+appY) and P28 (crtB), digested each with EdinbrickI using XbaI/PstI and set-up for ligations (L26 and L27). Stored overnight in the 16°C water bath.(HX)
