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From 2012.igem.org

Home Team Official Team Profile Project Parts Submitted Modeling Notebook Attributions

Contents 1 Notebook for week of July 2 to July 8 1.1 Design of Converter Plasmid 1.2 Design of Reporter Plasmid 1.3 Spinach Aptamer 1.4 CBB5 Caffeine Inducible Promoters 1.5 Inducible Odor

Notebook for week of July 2 to July 8

Design of Converter Plasmid

• Razan
  • 7/2 - Overlap PCR of Las Promoter and GFP+Terminator for control plasmid.
  • 7/5 - Amplification PCR of gel extracted band from overlap PCR.

Design of Reporter Plasmid

Spinach Aptamer

CBB5 Caffeine Inducible Promoters

• Peter
  • 7/2 - Performed PCR of BS_I20270 backbone for Gibson assembly
  • 7/3 - PCR purification on both PCRs. Gibson assembly of purified PCR. Gel Electrophoresis indicated that Gibson assembly was unsuccessful, as insert ORF 1 PCR was unsuccessful while vector PCR was successful. Erik independently performed PCR on the ORF 1. Used this, along with Dpn1 digested vector PCR to create a new Gibson.
  • 7/5 - Made new lab SOC under Aruko's supervision. Transformation of supposedly successful Gibson assembly
  • 7/6 - Unsuccessful transformation. Made a new second attempt.
  • 7/7 - Collected 3 successful small colonies from Gibson product assembly, as well as three obviously larger colonies from Gibson control plate which grew. Made cultures for each picked colony.

Inducible Odor