Edinburgh/7 August 2008

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<ul>
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<li><a href="https://2008.igem.org/Team:Edinburgh" title="Home"><span>Home</span></a></li>
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<li><a href="https://2008.igem.org/Team:Edinburgh/Project" title="Project" rel="dropmenu1_a"><span>The Project</span></a></li>
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<li><a href="https://2008.igem.org/Team:Edinburgh/Modeling" title="Modelling" rel="dropmenu2_a"><span>Modelling</span></a></li>
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<li><a href="https://2008.igem.org/Team:Edinburgh/Notebook" title="Notebook"><span>Notebook</span></a></li>
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<li><a href="https://2008.igem.org/Team:Edinburgh/Results" title="Results" rel="dropmenu1_a"><span>Results</span></a></li>
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<li><a href="http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2008&group=Edinburgh" title="Parts" rel="dropmenu1_a"><span>BioBrick Parts</span></a></li>
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<li><a href="https://2008.igem.org/Team:Edinburgh/Team" title="Team" ><span>The Team</span></a></li>
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<a href="https://2008.igem.org/Team:Edinburgh/Team">Overview</a>
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<a href="https://2008.igem.org/Team:Edinburgh/Team">Step1</a>
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<a href="https://2008.igem.org/Team:Edinburgh/Team">Step2</a>
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|-
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|align="left" width="150pt"|{{#calendar: title=Edinburgh |year=2008 | month=06}}
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|-
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|align="left" width="150pt"|{{#calendar: title=Edinburgh |year=2008 | month=07}}
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|-
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|align="left" width="150pt"|{{#calendar: title=Edinburgh |year=2008 | month=08}}
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|}
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:::: '''[[Edinburgh/6_August_2008|< Previous Entry]]'''
:::: '''[[Edinburgh/6_August_2008|< Previous Entry]]'''
Line 332: Line 6:
=== Thursday 7 August 08 ===
=== Thursday 7 August 08 ===
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* 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)
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* Subcultured from Plates 89-90 (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)
* 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)
+
* Ligation of rbs+''crtB'' (from M67) and pSB1A2+rbs+''crtI'' (from M50): rbs+''crtB'' as an insert cut with EcoRI/SpeI; rbs+''crtI''+pSB1A2 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.
+
* Double digestions performed: 2 of pSB1A2+rbs+''dxs'' (M72), pSB1A2+rbs+''crtE'' (M63) and pSB1A2+rbs+''lims1'' (from 0713536 in iGEM 07 box): pSB1A2+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)
+
* Attempted PCR of ''cenA'' ('''P52''') and ''cex'' ('''P53''') again using heat killed ''C. fimi'' as the template. The PCR products were checked on unnumbered gel, but failed again! (Yan, AM)
-
* PCR '''P54''' of pCstA. Run on '''gel 40'''. Results look promising (fairly prominent band <500bp). Purified. (CF)
+
* PCR '''P54''' of ''P<sub>cstA</sub>'' performed. 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 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)<br />
+
* '''Plates 96-97''' (glycogen assay) were spread with subcultures from Plate 43 (pSB1A2+rbs+''dxs''). Plate 96 = (-)glucose, plate 97 = (+)glucose. (HX)<br />
<br />
<br />
:::: '''[[Edinburgh/8_August_2008|Next Entry >]]'''
:::: '''[[Edinburgh/8_August_2008|Next Entry >]]'''

Latest revision as of 20:53, 29 October 2008

< Previous Entry

Week 8

Thursday 7 August 08

  • Subcultured from Plates 89-90 (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)
  • Ligation of rbs+crtB (from M67) and pSB1A2+rbs+crtI (from M50): rbs+crtB as an insert cut with EcoRI/SpeI; rbs+crtI+pSB1A2 as a vector cut with EcoRI/XbaI (L32). (Yan, AM)
  • Double digestions performed: 2 of pSB1A2+rbs+dxs (M72), pSB1A2+rbs+crtE (M63) and pSB1A2+rbs+lims1 (from 0713536 in iGEM 07 box): pSB1A2+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.
  • Attempted PCR of cenA (P52) and cex (P53) again using heat killed C. fimi as the template. The PCR products were checked on unnumbered gel, but failed again! (Yan, AM)
  • PCR P54 of PcstA performed. 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 96-97 (glycogen assay) were spread with subcultures from Plate 43 (pSB1A2+rbs+dxs). Plate 96 = (-)glucose, plate 97 = (+)glucose. (HX)


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