Edinburgh/11 August 2008

From 2008.igem.org

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<div id="colortab" class="ddcolortabs">
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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/Team" title="Team" ><span>The Team</span></a></li>
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<li><a href="http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2008&group=Edinburgh" title="Project" rel="dropmenu1_a"><span>BioBrick Parts</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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<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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=== Week 9 ===
=== Week 9 ===
==== Monday 11 August 08 ====
==== Monday 11 August 08 ====
-
* PCR products of P55 to P58 (''cenA'' and ''cex'') were run on '''Gel 41:''' ''cex'' with KOD polymerase (P57) generated a single band of the desired size. However, ''cex'' with Velocity polymerase and ''cenA'' with both KOD or Velocity polymerase failed. (YAN)
+
* PCR products of P55-P58 (''cenA'' and ''cex'') were run on '''Gel 41:''' ''cex'' with KOD polymerase (P57) generated a single band of the desired size. However, ''cex'' with Velocity polymerase and ''cenA'' with both KOD or Velocity polymerase failed. (Yan)
-
* PCR products P61 to P63 (''rrnB'', ''cenA'', ''cex'') were run on '''Gel 42'''. Very clear band for ''rrnB'', no bands for either ''cenA'' or ''cex''. (CF)
+
* PCR products P61-P63 (''rrnB'', ''cenA'', ''cex'') were run on '''Gel 42'''. Very clear band for ''rrnB'', no bands for either ''cenA'' or ''cex''. (CF)
-
* Made '''Plate 109/110''' (Transformation of L33: pBS1A2+pCstA), '''111/112''' (transformation of L34: pBS1A2+''dxs''+''crtE'') and Plate '''113/114''' (Transformation of L35: pBS1A2+''dxs''+''lims1''). (Yan)
+
* Made '''Plates 109-110''' (Transformation of L33: pBS1A2+''P<sub>cstA</sub>''), '''111-112''' (transformation of L34: pBS1A2+''dxs''+''crtE'') and '''113-114''' (Transformation of L35: pBS1A2+''dxs''+''lims1''). (Yan)
-
* Maxipreps '''X9''' of pSB1A2+rbs+''dxs'' (as for M72) and '''X10''' of pSB1A2P+rbs+''appY'' (as for M82). (AM)
+
* Made maxipreps '''X9''' of pSB1A2+rbs+''dxs'' (as for M72) and '''X10''' of pSB1A2P+rbs+''appY'' (as for M82). (AM)
* Redid PCR (KOD) of ''cex'' from ''C. fimi'' genomic DNA but with 10µl 50% glycerol, denaturing 1 min, annealing 60°C for 30s and extension 30s ('''P64'''). To be run on gel tomorrow. (AM)
* Redid PCR (KOD) of ''cex'' from ''C. fimi'' genomic DNA but with 10µl 50% glycerol, denaturing 1 min, annealing 60°C for 30s and extension 30s ('''P64'''). To be run on gel tomorrow. (AM)
* Digested P57 (''cex'' from KOD PCR) with EcoRI/PstI to confirm identity of the gene. ''cex'' has a PstI restriction site near its midpoint, so this digestion should generate two segments of approximately 750bp. To be run on gel tomorrow. (AM)
* Digested P57 (''cex'' from KOD PCR) with EcoRI/PstI to confirm identity of the gene. ''cex'' has a PstI restriction site near its midpoint, so this digestion should generate two segments of approximately 750bp. To be run on gel tomorrow. (AM)
* Digests performed using EcoRI/PstI to move ''crtE'' from BABEL2 (M79) to Edinbrick1. In each case, 3µl of vector was digested in 20µl total volume. The digests were run on a gel ready for excision, but gel showed no band corresponding to ''crtE'' (see '''gel 43''' showing a single band at ~3kb for BABEL2+''crtE'' and bands at ~2.5kb and 800bp for Edinbrick1). (AH)
* Digests performed using EcoRI/PstI to move ''crtE'' from BABEL2 (M79) to Edinbrick1. In each case, 3µl of vector was digested in 20µl total volume. The digests were run on a gel ready for excision, but gel showed no band corresponding to ''crtE'' (see '''gel 43''' showing a single band at ~3kb for BABEL2+''crtE'' and bands at ~2.5kb and 800bp for Edinbrick1). (AH)
-
* '''M109-M112''' Minipreps of pSB1A2+''crtB''+''crtI'', from plates 102. '''M113-M116''' Minipreps of pSB1A2+''glgC''-mut1,2 from plate 103. '''M117-M120''' Minipreps of pSB1A2+''glgC''-mut1,2,3 from plate 94 (OG)
+
* Minipreps '''M109-M112''' (putative pSB1A2+''crtB''+''crtI'', plate 102), '''M113-M116''' (pSB1A2+''glgC''-mut1,2, plate 103) and '''M117-M120''' (pSB1A2+''glgC''-mut1,2,3, plate 94) were made. (OG)
-
* Double digestion of M109 to M120 with EcoRI/PstI, run on '''gel 44'''. M110-102 (pSB1A2+''crtB''+''crtI'') have a band at 2.5kb as expected. M109 (pSB1A2+''crtB''+''crtI'') gave a band at 3kb instead, so should be ignored from now on. (Yan)
+
* Double digests of M109-M120 with EcoRI/PstI were run on '''gel 44'''. M110-M112 (pSB1A2+''crtB''+''crtI'') have a band at 2.5kb as expected. M109 (pSB1A2+''crtB''+''crtI'') gave a band at 3kb instead, so should be ignored from now on. (Yan)
-
* Subbed white colonies from plates 105 (pSB1A2+''crtY'') and 106 (pSB1A2+rbs+''crtY'') to '''Plate 108'''. (CF)
+
* Subbed white colonies from plates 105 (pSB1A2+''crtY'') and 106 (pSB1A2+rbs+''crtY'') to '''Plate 108'''. (CF)<br />
 +
<br />
 +
 
 +
:::: '''[[Edinburgh/12_August_2008|Next Entry >]]'''

Latest revision as of 20:53, 29 October 2008

< Previous Entry

Week 9

Monday 11 August 08

  • PCR products of P55-P58 (cenA and cex) were run on Gel 41: cex with KOD polymerase (P57) generated a single band of the desired size. However, cex with Velocity polymerase and cenA with both KOD or Velocity polymerase failed. (Yan)
  • PCR products P61-P63 (rrnB, cenA, cex) were run on Gel 42. Very clear band for rrnB, no bands for either cenA or cex. (CF)
  • Made Plates 109-110 (Transformation of L33: pBS1A2+PcstA), 111-112 (transformation of L34: pBS1A2+dxs+crtE) and 113-114 (Transformation of L35: pBS1A2+dxs+lims1). (Yan)
  • Made maxipreps X9 of pSB1A2+rbs+dxs (as for M72) and X10 of pSB1A2P+rbs+appY (as for M82). (AM)
  • Redid PCR (KOD) of cex from C. fimi genomic DNA but with 10µl 50% glycerol, denaturing 1 min, annealing 60°C for 30s and extension 30s (P64). To be run on gel tomorrow. (AM)
  • Digested P57 (cex from KOD PCR) with EcoRI/PstI to confirm identity of the gene. cex has a PstI restriction site near its midpoint, so this digestion should generate two segments of approximately 750bp. To be run on gel tomorrow. (AM)
  • Digests performed using EcoRI/PstI to move crtE from BABEL2 (M79) to Edinbrick1. In each case, 3µl of vector was digested in 20µl total volume. The digests were run on a gel ready for excision, but gel showed no band corresponding to crtE (see gel 43 showing a single band at ~3kb for BABEL2+crtE and bands at ~2.5kb and 800bp for Edinbrick1). (AH)
  • Minipreps M109-M112 (putative pSB1A2+crtB+crtI, plate 102), M113-M116 (pSB1A2+glgC-mut1,2, plate 103) and M117-M120 (pSB1A2+glgC-mut1,2,3, plate 94) were made. (OG)
  • Double digests of M109-M120 with EcoRI/PstI were run on gel 44. M110-M112 (pSB1A2+crtB+crtI) have a band at 2.5kb as expected. M109 (pSB1A2+crtB+crtI) gave a band at 3kb instead, so should be ignored from now on. (Yan)
  • Subbed white colonies from plates 105 (pSB1A2+crtY) and 106 (pSB1A2+rbs+crtY) to Plate 108. (CF)


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