Team:Osaka/Notebook

Weekly Summaries

Week 1

Went through basic molecular biology lab skills and techniques with the new members. Extracted most of the existing parts required for our project from the iGEM 2012 Spring distribution plates, transformed and amplified the DNA for future use.DNA extraction & purification (miniprep) of transformed parts, & gel runs to confirm lengths. Ligations to put together some common combinations (eg promoter + RBS).

We transfered redioresistance parts from one vector(pSB1C3) to another vector(pSB1A3) and transformed plasmid DNA into Rosetta E.coli.

Week 2

We run the SDS-PAGE gel to check the expression of the gene products at the protein level. The result of DNA sequencing demonstrated that there is a mutation in PprA(Silent mutation).

Week 3

Extraction & amplification of more basic parts from the distribution plates, plus ligation of a few more parts. Not much progress due to lab being closed for Obon holidays.

Completion of radioresistance parts and drafting of damage tolerance assay protocol.

Week 4

We were going to assemble a promoter evaluation device utilizing firefly and renilla luciferase, but did not make much progress due to low transformation efficiency(?).

Week 5

Preliminary test (damage tolerance, Mitomycin C).

We were going to assemble a promoter evaluation device utilizing firefly and renilla luciferase, but did not make much progress due to low transformation efficiency(?).

Week 6

Damage tolerance assay (Mitomycin C): transformed cells were exposed to DNA damaging agents and then incubated on agar plates, and viability relative to control used as an evaluation of the abilities of PprI, PprA, PprM, RecA to confer resistance to radiation-induced DNA damage

Week 7

Damage tolerance assay (Mitomycin C)

Preliminary test (damage tolerance, Hydrogen peroxide)

Week 8

Damage tolerance assay (Hydrogen peroxide)

Week 9

Damage tolerance assay (Hydrogen peroxide)

Logbook Entries

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