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Things we did today

Wetlab work

Margaret

5 colonies of rep, P1 lytic region, and aadA(pRL1383a) region, each. pSB1A3 was used as a control.

PCR verification of aadA from pRL1383a using wrong conditions.

I changed the annealing time to 3min 30 seconds instead of the extension time, so the sites were not amplified properly. I ran the aadA(pRL1383a) construct on a gel and got a few bands, none of which were the right size. One of them should have been right though, so I am assuming this ligation did not work.
I ran another PCR verification on rep, P1, and the omega interposon and afterwards a gel.

PCR verification of rep, P1 lytic regbion, and omega interposon ligation from 8-13

Margaret

Plasmid preps from yesterday did not yield very high quantities. I am setting up the culture for oriV1-4 and aadA(BB) 4 so I can do another plasmid prep tomorrow.

Margaret

digest of I14032 with SpeI and PstI. I want to use this to ligate to the aadA(BB) construct.

Margaret

Verified sequence of oriT, inserted into pSB1A2. The sequences of oriT and the BioBrick sites are correct.

Margaret & Grace

We are having trouble with TT, RBS and Promoter ligations, so we decided to synthesize these parts.
B0016, I14032, B0030, & B0034

History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson

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