Edinburgh/7 August 2008

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(→Thursday 7 August 08)
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=== 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)
+
* 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 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 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 >]]'''

Revision as of 11:37, 28 August 2008

Edinburgh iGEM 2008

 

June
MTWTFSS
            [http://2008.igem.org/wiki/index.php?title=Edinburgh/1_June_2008&action=edit 1]
[http://2008.igem.org/wiki/index.php?title=Edinburgh/2_June_2008&action=edit 2] [http://2008.igem.org/wiki/index.php?title=Edinburgh/3_June_2008&action=edit 3] [http://2008.igem.org/wiki/index.php?title=Edinburgh/4_June_2008&action=edit 4] [http://2008.igem.org/wiki/index.php?title=Edinburgh/5_June_2008&action=edit 5] [http://2008.igem.org/wiki/index.php?title=Edinburgh/6_June_2008&action=edit 6] [http://2008.igem.org/wiki/index.php?title=Edinburgh/7_June_2008&action=edit 7] [http://2008.igem.org/wiki/index.php?title=Edinburgh/8_June_2008&action=edit 8]
[http://2008.igem.org/wiki/index.php?title=Edinburgh/9_June_2008&action=edit 9] [http://2008.igem.org/wiki/index.php?title=Edinburgh/10_June_2008&action=edit 10] [http://2008.igem.org/wiki/index.php?title=Edinburgh/11_June_2008&action=edit 11] [http://2008.igem.org/wiki/index.php?title=Edinburgh/12_June_2008&action=edit 12] [http://2008.igem.org/wiki/index.php?title=Edinburgh/13_June_2008&action=edit 13] [http://2008.igem.org/wiki/index.php?title=Edinburgh/14_June_2008&action=edit 14] [http://2008.igem.org/wiki/index.php?title=Edinburgh/15_June_2008&action=edit 15]
[http://2008.igem.org/wiki/index.php?title=Edinburgh/16_June_2008&action=edit 16] [http://2008.igem.org/Edinburgh/17_June_2008 17] [http://2008.igem.org/wiki/index.php?title=Edinburgh/18_June_2008&action=edit 18] [http://2008.igem.org/Edinburgh/19_June_2008 19] [http://2008.igem.org/Edinburgh/20_June_2008 20] [http://2008.igem.org/wiki/index.php?title=Edinburgh/21_June_2008&action=edit 21] [http://2008.igem.org/wiki/index.php?title=Edinburgh/22_June_2008&action=edit 22]
[http://2008.igem.org/wiki/index.php?title=Edinburgh/23_June_2008&action=edit 23] [http://2008.igem.org/Edinburgh/24_June_2008 24] [http://2008.igem.org/Edinburgh/25_June_2008 25] [http://2008.igem.org/Edinburgh/26_June_2008 26] [http://2008.igem.org/Edinburgh/27_June_2008 27] [http://2008.igem.org/Edinburgh/28_June_2008 28] [http://2008.igem.org/Edinburgh/29_June_2008 29]
[http://2008.igem.org/Edinburgh/30_June_2008 30]
July
MTWTFSS
  [http://2008.igem.org/Edinburgh/1_July_2008 1] [http://2008.igem.org/Edinburgh/2_July_2008 2] [http://2008.igem.org/Edinburgh/3_July_2008 3] [http://2008.igem.org/Edinburgh/4_July_2008 4] [http://2008.igem.org/Edinburgh/5_July_2008 5] [http://2008.igem.org/Edinburgh/6_July_2008 6]
[http://2008.igem.org/Edinburgh/7_July_2008 7] [http://2008.igem.org/Edinburgh/8_July_2008 8] [http://2008.igem.org/Edinburgh/9_July_2008 9] [http://2008.igem.org/Edinburgh/10_July_2008 10] [http://2008.igem.org/Edinburgh/11_July_2008 11] [http://2008.igem.org/Edinburgh/12_July_2008 12] [http://2008.igem.org/Edinburgh/13_July_2008 13]
[http://2008.igem.org/Edinburgh/14_July_2008 14] [http://2008.igem.org/Edinburgh/15_July_2008 15] [http://2008.igem.org/Edinburgh/16_July_2008 16] [http://2008.igem.org/Edinburgh/17_July_2008 17] [http://2008.igem.org/Edinburgh/18_July_2008 18] [http://2008.igem.org/Edinburgh/19_July_2008 19] [http://2008.igem.org/Edinburgh/20_July_2008 20]
[http://2008.igem.org/Edinburgh/21_July_2008 21] [http://2008.igem.org/Edinburgh/22_July_2008 22] [http://2008.igem.org/Edinburgh/23_July_2008 23] [http://2008.igem.org/Edinburgh/24_July_2008 24] [http://2008.igem.org/Edinburgh/25_July_2008 25] [http://2008.igem.org/Edinburgh/26_July_2008 26] [http://2008.igem.org/Edinburgh/27_July_2008 27]
[http://2008.igem.org/Edinburgh/28_July_2008 28] [http://2008.igem.org/Edinburgh/29_July_2008 29] [http://2008.igem.org/Edinburgh/30_July_2008 30] [http://2008.igem.org/Edinburgh/31_July_2008 31]
August
MTWTFSS
        [http://2008.igem.org/Edinburgh/1_August_2008 1] [http://2008.igem.org/wiki/index.php?title=Edinburgh/2_August_2008&action=edit 2] [http://2008.igem.org/Edinburgh/3_August_2008 3]
[http://2008.igem.org/Edinburgh/4_August_2008 4] [http://2008.igem.org/Edinburgh/5_August_2008 5] [http://2008.igem.org/Edinburgh/6_August_2008 6] [http://2008.igem.org/Edinburgh/7_August_2008 7] [http://2008.igem.org/Edinburgh/8_August_2008 8] [http://2008.igem.org/Edinburgh/9_August_2008 9] [http://2008.igem.org/Edinburgh/10_August_2008 10]
[http://2008.igem.org/Edinburgh/11_August_2008 11] [http://2008.igem.org/Edinburgh/12_August_2008 12] [http://2008.igem.org/Edinburgh/13_August_2008 13] [http://2008.igem.org/Edinburgh/14_August_2008 14] [http://2008.igem.org/Edinburgh/15_August_2008 15] [http://2008.igem.org/Edinburgh/16_August_2008 16] [http://2008.igem.org/Edinburgh/17_August_2008 17]
[http://2008.igem.org/Edinburgh/18_August_2008 18] [http://2008.igem.org/Edinburgh/19_August_2008 19] [http://2008.igem.org/Edinburgh/20_August_2008 20] [http://2008.igem.org/Edinburgh/21_August_2008 21] [http://2008.igem.org/Edinburgh/22_August_2008 22] [http://2008.igem.org/wiki/index.php?title=Edinburgh/23_August_2008&action=edit 23] [http://2008.igem.org/wiki/index.php?title=Edinburgh/24_August_2008&action=edit 24]
[http://2008.igem.org/Edinburgh/25_August_2008 25] [http://2008.igem.org/Edinburgh/26_August_2008 26] [http://2008.igem.org/Edinburgh/27_August_2008 27] [http://2008.igem.org/Edinburgh/28_August_2008 28] [http://2008.igem.org/Edinburgh/29_August_2008 29] [http://2008.igem.org/wiki/index.php?title=Edinburgh/30_August_2008&action=edit 30] [http://2008.igem.org/wiki/index.php?title=Edinburgh/31_August_2008&action=edit 31]
< 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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