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Contents 1 Team Cinnamaldehyde & Vanillin 1.1 June 28th 1.1.1 Transformation of 4-coumarate-CoA Ligase (4CL) and caffeic acid-O-methyl transferase (COMT) 1.1.2 Rhodobacter sphaeroides Inoculation 1.2 July 30th 1.2.1 Miniprep 4CL 1.3 July 4th 1.3.1 PCR of 4-CL

Team Cinnamaldehyde & Vanillin

Research Designs and Methods:

The goal for cloning is to place each gene in the cinnamaldehyde pathways under a constitutive pTET and arabinose promoter pBAD for future characterization of each part. Our final construct should consist of a single pSB1C3 plasmid with all the genes inserted along with its ribosomal binding site and terminator:

June 28th

Transformation of 4-coumarate-CoA Ligase (4CL) and caffeic acid-O-methyl transferase (COMT)

Experimenter: Joe

Aim: Transform biobricks 4CL ( 2013 Distribution Kit, Plate 2, Well 17P) and COMT (2013 Distribution Kit, Plate 5, Well 19D) for miniprep and later amplification through PCR

Results: Transformation done according transformation procedures. Colonies grown overnight at 37C. Only 4-CL grew overnight. No colonies were observed for COMT. Inoculated a colony of 4CL for miniprep

Rhodobacter sphaeroides Inoculation

Experimenter: Joe

Aim: A colony of Rhodobacter sphaeroides was inoculated in 5mL LB culture for DNA extraction

July 30th

Miniprep 4CL

Experimenter: Joe

Aim: Miniprep 5mL innoculation of 4CL with Qiagen Miniprep Kit

July 4th

PCR of 4-CL

Experimenter: Joe

Aim: PCR to amplify 4-CL from biobrick with biobrick VF2 and VR primers to confirm the size of 4-CL

Results: Multiple bands w