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CAROL
Preparing Competent Cells
- Transformed plasmid, pCS26 into Top 10 cells with NEB Quick Ligase Buffer and Anarctic Phosphatase Buffer to verify if the E.coli cells are able to uptake the larger plasmid. We wanted to check the transformation efficiency of a 10KB plasmid. This was plated on LB+Kan plates.
- To prepare for making competent cells, we grew overnight cultures in 5 mL of LB broth of Top 10 cells and XL Gold Ultracompotent cells (Stratagene)
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CHINMOYEE
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EMILY
An Assortment of Miscellaneous Tasks
This morning I helped Carol and Jeremy work with Johnathon, Carol's high school student. We made LB+ CM 35 broth as well as AK, AC, C and K plates. We also gave him a tour (led by Jeremy) of the Health Sciences Centre and our labs. In the afternoon I read through the Ethics Webinar report online and did a write-up on the Ethics Webinar for Jamie for the July newsletter. I also edited some write-ups for Mandi that are going up on the Wiki to show our collaboration with both the U of A and U of L teams. Sequencing came back today and it looks like the B0015 terminator was not successfully cloned in. Because of this, tomorrow I will have to re-digest J13002-LuxOD47E and B0015 to try construction again. I will follow this with transformation and plating.
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FAHD
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IMAN
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JAMIE
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JEREMY
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KATIE
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KEVIN
1. Plasmid Isolation of B0034
The overnights of B0034 were made for me by Thane on Sunday. Thank you Thane. I have isolated the plasmids from these overnight cultures, and the purity and concentration of them were measured using nanodrop. They were pure enough to move on with.
2. Construction of Pqrr4B0034
Since our Luciferase did not turn out great, we turned our attention to GFP:LVA. But because the part with GFP:LVA does not contain RBS, I need to constrct Pqrr4+B0034 first. Pqrr4 was cut with SpeI+PstI and B0034 was cut with XbaI+PstI. Pqrr4 was then phosphotase treated, and the two were ligated together. The ligated product was finally transformed into TOP10 cells.
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MANDY
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PATRICK
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PRIMA
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STEFAN
Meeting at Christian's Lab
Today was spent preparing the synthetic kingdom for the meeting at
ICT with an educator friend of Sonja's. Mandy and I also showed the high school student that's with us for the week around second
life. At the meeting we got quite a bit of insight on how to further
improve our island.
For my portion:
- Have "exhibits"
- Have a narrative (why do we care?)
- Exploration should be more guided
- more engineered cells
- Tell people about what they are seeing
- Integrate tutorial
- Squid buddy that shows you around
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VICKI
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