Edinburgh/1 August 2008


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Friday 1 August 08

 * Primers for pCstA ordered. (CF)
 * White colonies from Thursday's transformations were subplated as follows:
 * Plate 76 (L26: pSB1A2+rbs+appY) to Plate 83
 * Plate 77 (L27: pSB1A2+rbs+crtB) to Plate 84
 * Plate 80 (L28: pSB1A2+cenA) to Plate 85
 * Plates 81/82 (L29: pSB1A2+cex) to Plate 86. (AM/HX/OG)
 * First digestion: M68-M71 (pSB1A2+glgC-mut1,2) with EcoRI/PstI, M72-M75 (pSB1A2+rbs+dxs) with EcoRI, M76-M79 (BABEL2+crtE) with EcoRI/PstI. Run on Gel 33. Results: M68, 69, 71: vector band around 2 kb, no glgC band; M72, 73, 74: single band around 4 kb, expected length for vector+gene; M76-79: crtE band <1 kb, vector band ~2 kb. It is not clear into which vector crtE (M76-79) is inserted, so sequencing will be necessary. (AM)
 * Second digestion: M68 and M69 with a) EcoRI, b) EcoRI/PstI; M72 with a) EcoRI b) SacI/SpeI. Run on Gel 34. Results: Single and double digests of M68 and M69 yielded similar results, hence pSB1A2+glgC-mut1,2 ligation or transformation failed; digests of M72 yielded expected bands, hence pSB1A2+rbs+dxs probably successful. (AM)
 * P36 (glgC-mut1,2 excision from BABEL2) and L25 (P36 + Edinbrick1) were run again on Gel 35. Results: P36 viewed at lower illumination yields one band of the desired length (1.5kb); L25 yields two bands around 1.5kb, a thick band at 3kb and several fainter bands including one at 2kb (length of pSB1A2). (AM)


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