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Team:Arizona State/Notebook
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| - | <h2>Week 3</h2> | + | <h2>June: Week 3</h2> |
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<li>Started making amp plates in preparation for experimentation</li> | <li>Started making amp plates in preparation for experimentation</li> | ||
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| - | <h2>Week 7</h2> | + | <h2>July: Week 7</h2> |
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<li>Performed restriction digest of pCadA, pBN, and YebF</li> | <li>Performed restriction digest of pCadA, pBN, and YebF</li> | ||
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| - | <h2>Week 11</h2> | + | <h2>August: Week 11</h2> |
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<li>Grew up six cultures of ligations</li> | <li>Grew up six cultures of ligations</li> | ||
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| - | <h2>Week | + | <h2>September: Week 15</h2> |
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<li>Made ligations of FLU+1c3, MelA+1c3, Flu+J04..., MelA+J04...</li> | <li>Made ligations of FLU+1c3, MelA+1c3, Flu+J04..., MelA+J04...</li> | ||
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
