Team:Arizona State/Notebook

From 2013.igem.org

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</body>
</body>
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<h1>May: Week 1</h1>
+
<h2>May: Week 1</h2>
<ul>
<ul>
<li>First ASU iGEM 2013 with new team members</li>
<li>First ASU iGEM 2013 with new team members</li>
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</ul>
</ul>
-
<h2>Week 3</h2>
+
<h2>June: Week 3</h2>
 +
<ul>
 +
<li>Started making amp plates in preparation for experimentation</li>
 +
</ul>
<h2>Week 4</h2>
<h2>Week 4</h2>
 +
<ul>
 +
<li>Started growing up colonies of E. coli K12 MG1655, BL21 (DE3),and N10 B strains along with glycerol stocks</li>
 +
<li>Performed a restriction, ligation and transformation of the three plasmids and grew them up on a plate</li>
 +
<li>No colony growth of any of the plasmids</li>
 +
</ul>
-
<h2>June: Week 5</h2>
+
<h2>Week 5</h2>
 +
<ul>
 +
<li>Performed a PCR amplification of the cadA promoter along with a gel screen that found the band at the appropriate band length</li>
 +
<li>Transformed and plated both J61100 and E1010 with no growth on either plates</li>
 +
<li>Gel screen of yebF without the stop codon never showed up on the gel. Ran transformation again.</li>
 +
<li>Restricted and ligated cadA and BN promoters</li>
 +
</ul>
 +
 
 +
<h2>Week 6</h2>
 +
<ul>
 +
<li>The gel screen of yebF without the stop codon was finally successful</li>
 +
<li>Transformation of INPNC-MCS and RFP control onto N10B cells was successful</li>
 +
<li>No growth from transformation of E0020, E0030, and E0040</li>
 +
<li>Created liquid cultures of E0240 and pCadA in LB Chloro.</li>
 +
</ul>
 +
 
 +
<h2>July: Week 7</h2>
 +
<ul>
 +
<li>Performed restriction digest of pCadA, pBN, and YebF</li>
 +
<li>Performed seven different ligations</li>
 +
<li>Miniprepped and nanodropped E0240 and pCadA.</li>
 +
<li>Miniprepped YebF+GMCSF and PelB leader sequence.</li>
 +
<li>Flow cytometry indicated that there was no GFP</li>
 +
<li>Transformed J23100 yet no plate growth</li>
 +
</ul>
 +
 
 +
<h2>Week 8</h2>
 +
<ul>
 +
<li>Ordered Antigen primers</li>
 +
<li>Ligation and transformation of PelB+GMCSF+pSB1C3/4K5</li>
 +
<li>Miniprep showed no successful ligation and transformation</li>
 +
</ul>
 +
 
 +
<h2>Week 9</h2>
 +
<ul>
 +
<li>PCR of MelanA and Flu M1 worked via confirmation from gel screen</li>
 +
<li>Nissle would not grow</li>
 +
<li>Transformed I13522 into N10B on LB Amp</li>
 +
</ul>
 +
 
 +
<h2>August: Week 11</h2>
 +
<ul>
 +
<li>Grew up six cultures of ligations</li>
 +
<li>Miniprepped the PSB1c3 backbone and grew up additional backbone in cultures</li>
 +
<li>Transformed the GFP and GMCSF in the backbone</li>
 +
<li>Ran LLO pcr on gel and all had correct band lengths</li>
 +
</ul>
 +
 
 +
<h2>September: Week 15</h2>
 +
<ul>
 +
<li>Made ligations of FLU+1c3, MelA+1c3, Flu+J04..., MelA+J04...</li>
 +
<li>Made liquid cultures of non-red colonies from FLU+1c3, MelA+1c3, Flu+J04..., MelA+J04 transformation</li>
 +
<li>Biobricks made and sent: LLO with promoter RBS, YEb-GMCSF, GMCSF, FLU1, and MelanA</li>
 +
</ul>
</html>
</html>

Latest revision as of 23:31, 27 September 2013

May: Week 1

  • First ASU iGEM 2013 with new team members
  • Introduction to the competition, registration logistics, summer schedules
  • Idea Brainstorming
  • Review of biobrick cloning, Golden Gate assembly, safety training

Week 2

  • Idea Brainstorming--narrowed down to two project ideas

Safety Training:

  • Biosafety and Bloodborne Pathogens
  • Lab Safety
  • Fire Safety
  • Recombinant DNA Safety
  • Hazardous Waste Management

June: Week 3

  • Started making amp plates in preparation for experimentation

Week 4

  • Started growing up colonies of E. coli K12 MG1655, BL21 (DE3),and N10 B strains along with glycerol stocks
  • Performed a restriction, ligation and transformation of the three plasmids and grew them up on a plate
  • No colony growth of any of the plasmids

Week 5

  • Performed a PCR amplification of the cadA promoter along with a gel screen that found the band at the appropriate band length
  • Transformed and plated both J61100 and E1010 with no growth on either plates
  • Gel screen of yebF without the stop codon never showed up on the gel. Ran transformation again.
  • Restricted and ligated cadA and BN promoters

Week 6

  • The gel screen of yebF without the stop codon was finally successful
  • Transformation of INPNC-MCS and RFP control onto N10B cells was successful
  • No growth from transformation of E0020, E0030, and E0040
  • Created liquid cultures of E0240 and pCadA in LB Chloro.

July: Week 7

  • Performed restriction digest of pCadA, pBN, and YebF
  • Performed seven different ligations
  • Miniprepped and nanodropped E0240 and pCadA.
  • Miniprepped YebF+GMCSF and PelB leader sequence.
  • Flow cytometry indicated that there was no GFP
  • Transformed J23100 yet no plate growth

Week 8

  • Ordered Antigen primers
  • Ligation and transformation of PelB+GMCSF+pSB1C3/4K5
  • Miniprep showed no successful ligation and transformation

Week 9

  • PCR of MelanA and Flu M1 worked via confirmation from gel screen
  • Nissle would not grow
  • Transformed I13522 into N10B on LB Amp

August: Week 11

  • Grew up six cultures of ligations
  • Miniprepped the PSB1c3 backbone and grew up additional backbone in cultures
  • Transformed the GFP and GMCSF in the backbone
  • Ran LLO pcr on gel and all had correct band lengths

September: Week 15

  • Made ligations of FLU+1c3, MelA+1c3, Flu+J04..., MelA+J04...
  • Made liquid cultures of non-red colonies from FLU+1c3, MelA+1c3, Flu+J04..., MelA+J04 transformation
  • Biobricks made and sent: LLO with promoter RBS, YEb-GMCSF, GMCSF, FLU1, and MelanA

Retrieved from "http://2013.igem.org/Team:Arizona_State/Notebook"