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Edinburgh/6 August 2008
From 2008.igem.org
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Week 8
Wednesday 6 August 08
- Primers have arrived for pCstA. PCR carried out using E. coli cells as the template (P51). This was unsuccessful. (CF)
- Digestion of M67 (pSB1A2+rbs+crtB) with EcoRI/SpeI and M50 (pSB1A2+rbs+crtI) with EcoRI/XbaI in preparation for combining the two, with the former as insert and the latter as vector. (CF)
- Re-digestion of M97-M99 and M101 (all pSB1A2+cenA) and M105 and M108 (pSB1A2+cex) with a) EcoRI, b) EcoRI/PstI, and M92 (pSB1A2+cex) with a) PstI, b) EcoRI/PstI. All run on gel 38. (Yan)
- Transformation of L30 (Self-ligation of BABEL2+glgC-mut1,2,3) to plates 89-90) and L31 (glgC-mut1,2 to Edinbrick1) to plates 91-92. (HX)
- Maxipreps X6 (as M50, pSB1A2+rbs+crtI) and X7 (as M63: pSB1A2-+rbs+crtE) made. (AM)
- Analytical digests using a) EcoRI, b) EcoRI/PstI of M70-M71 (pSB1A2+glgC-mut1,2) and M92 (pSB1A2+cex). Run on gel 39. Results do not look good. Bands are wrong size to correspond to vector and no bands match an expected insert size. (CF)
- Colony on plate 56 (M67, pSB1A2+rbs+crtB) subbed to plate 93. (CF)
- Overnight culture made from plate 93 (pSB1A2+rbs+crtB) ready for maxiprep tomorrow. (AM, AH)
