Team:WashU StLouis/Notebook
Notebook
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Micah, Maddie, Alex
Lab Work
- Prepared LB Media
- Re-suspended DNA (D. Radiodurans Uracil DNA Glycosylate 2) from iGEM kit
- Transformed Uracil DNA Glycosylate 2 in psB1C3 plasmid into MHD42 competent cells (Practice transformation)
- Prepared LB Agar
- Prepared overnight culture to make competent cells (MHD42)
Outside Work / Discussion
Micah, Maddie
Lab Work
- Prepared MHD42 competent cells
- Checked on transformed MHD42 cells with Uracil DNA Glycosylate 2
- Transformation was a success with 100 colonies
- Plate was stored in 4 deg. Celsius for future use
- Prepared TSS Buffer
Outside Work / Discussion
No Work!
No Work!
Micah, Maddie, Alex, Mark, Collin
Lab Work
- Prepared LB + Chloramphenicol (CM) plates
- Prepared Ampicillin solution to be used for plates in the future
- Made more LB Agar
Outside Work / Discussion
- Discussed shifting focus from Dsup gene to a variety of UV radiation resistance gene
- Found more genes to compare to Dsup (i.e. phrA in tardigrades and cyanobacteria)
- Met with Dr. Brennan to discuss updates
Micah, Maddie, Alex, Mark, Collin
Lab Work
- Prepared DH5α competent cells
- Tested MHD42 cells for competency
- Tested DH5α cells for competency
Outside Work / Discussion
- Fleshed out project and decided specific parts to transform [BBa_B0034 (RBS), BBa_R0010 (lac promoter), and BBa_K1499200 (uvsE gene)] into MHD42 cells during the next day
- The Deinococcus radiodurans UV DNA damage endonuclease (uvsE) gene is a UV radiation damage repair gene from the Stanford-Brown-Spelman 2014 team. We plan to use this to compare to other UV radiation repair genes.
- Planned out more long-term goals such as collaborations with other teams (especially those in the Midwest)
Lab Work
- Checked competency test for MHD42 cells
- Test was successful!
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- Checked competency test for DH5α
- Test showed that the cells were competent but had a low transformation efficiency
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- Prepared LB + Ampicillin (Amp) plates
- Resuspeded parts BBa_B0034, BBa_R0010, and BBa_K1499200 from current and past distribution kits
- Transformed part BBa_B0034 (with plasmid psB1A2) into MHD42 competent cells
- Transformed part BBa_R0010 (with plasmid psB1C3) into MHD42 competent cells
- Transformed part BBa_K1499200 (with plasmid psB1C3) into MHD42 competent cells
- Prepared overnight culture of DH5α competent cells
Outside Work / Discussion
- Discussed modelling ideas
- Learned how to design primers from Eugene
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