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From 2013.igem.org

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

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.

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

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

Week 13

• 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