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Team:British Columbia/Notebook/Flavours
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| + | 1.2 Research Designs and Methods | ||
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| + | 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'''== | =='''June 28th'''== | ||
Revision as of 22:09, 31 August 2013
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Team Cinnamaldehyde & Vanillin
1.2 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
