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Team:UTK-Knoxville/21 June 2013
From 2013.igem.org
June 21st, 2013
Eric is going to go ahead and transform the plasmid into chemi comp knockout strains, as well as digest the plasmid to confirm it. We will begin characterization when that is complete.
We didn’t amplify HemAT from the correct gDNA. The drive said that it was from MG1655, but I have fixed it. I got the cells(R. Leg) from Eric, and I will prep the media they need to be grown on when he emails me the protocol.
I got the R. Leg strain from Eric which contains the HemAT gene. I have made the TYC media to grow R. Leg in to extract gDNA. Eric recommended 48-72 hours to grow, so I will leave them for the weekend and extract/PCR gDNA Monday.
TYC Media(1 liter)
5g tryptone
3g yeast extract
.5g CaCl2
We didn’t amplify RcoM from the correct gDNA. The drive file said that it was from MG1655, but I have fixed it. I got the cells from Eric, and I will prep the media they need to be grown on when he emails me the protocol.
Eric brought TY liquid media for the Azo strain that contains RcoM, cells are growing in liquid media now and the Azo plate is in the cold room. Eric recommended 48-72 hours for them to grow before I extracted gDNA, so I will leave them for the weekend.
TY media(1 liter)
5g yeast extract
10g tryptone
