Edinburgh/7 August 2008

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Revision as of 11:30, 20 August 2008

Edinburgh iGEM 2008

 

Week 8

Thursday 7 August 08

  • Subcultured from Plates 89 and 90 (BABEL2+glgC-mut1,2,3) to Plate 94. Plate 91 (pSB1A2+glgC-mut1,2) showed no growth, so was discarded. Plate 92 (pSB1A2+glgC-mut1,2) had a blue smear and two white colonies. These colonies were subbed to Plate 95. (HX)
  • Maxiprep X8 (as M67: pSB1A2+rbs+crtB) made. Slight hiccup after step 6. (Cold solution 3 was added and incubation on ice was skipped. Centrifugation continued for 2 minutes before mistake was realised, after which the solution was mixed and incubated on ice for ten minutes (continuing with step 7). (AM, AH)
  • Combination (Ligation) of rbs+crtB (from M67) and rbs+crtI (from M50): rbs+crtB as an insert cut with EcoRI/SpeI; rbs+crtI as a vector cut with EcoRI/XbaI (L32). (Yan, AM)
  • Double digestion of three biobricks: 2 of rbs+dxs (from M72), rbs+crtE (from M63) and rbs+lims1 (from 0713536 in iGEM 07 box): rbs+dxs as a vector cut with SpeI/PstI; rbs+crtE as an insert cut with XbaI/PstI; rbs+lims1 as an insert cut with XbaI/PstI.
  • Made PCR of cenA and cex again, stored in P52 (cenA) and P53 (cex). The PCR products were checked on gel, but failed again! (Yan, AM)
  • PCR P54 of pCstA. Run on gel 40. Results look promising (fairly prominent band <500bp). Purified. (CF)
  • Plates made for testing glycogen assay. 2x LB agar, ampicillin; 2x LB agar, ampicillin, 2% glucose ready for spreading with E. coli. (SK)
  • Plates 96 and 97 (glycogen assay) were spread with subcultures from Plate 43. Plate 96 = (-)glucose, plate 97 = (+)glucose. (HX)
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