Edinburgh/6 August 2008

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Edinburgh iGEM 2008

L31 ligation of P36 (glgC-mut1,2, hopefully, could be P39) to Edinbrick1 completed. (OG)
Primers have arrived for pCstA. Chris carried out the PCR (that would be P51). 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)
- Not enough time to purify/ligate due to high demand for agarose gel following lunchtime. Shall do tomorrow morning at 9. (AM)
Re-digestion of M97, M98, 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 (because gel 37 did not look good). (Yan)
Transformation of L30 (Self-ligation of BABEL2+glgC-mut1,2,3) (plates 89 and 90) and L31 (glgC-mut1,2 to Edinbrick1) (plates 91 and 92) onto B/W plates. (HX)
Maxipreps X6 (as M50, pSB1A2+rbs+crtI) and X7 (as M63: pSB1A2-+rbs+crtE) made. (AM)
Overnight culture made from plate 93 (pSB1A2+rbs+crtB) ready for maxiprep tomorrow. (AM, AH)
Analytical digests using a) EcoRI, b) EcoRI/PstI of M70, M71 (both 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)