Notebook
Click on a date to see what we did on that day! Click the double arrowheads to switch months.
June 2017 | ||||||
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June 1
Present: Micah, Maddie, Alex
Lab Work
- Prepared LB Media [Micah, Maddie, Alex]
- Re-suspended DNA (D. Radiodurans Uracil DNA Glycosylate 2) from iGEM kit [Micah, Maddie, Alex]
- Transformed Uracil DNA Glycosylate 2 in psB1C3 plasmid into MHD42 competent cells (Practice transformation) [Micah, Maddie, Alex]
- Prepared LB Agar [Micah, Maddie, Alex]
- Prepared overnight culture to make competent cells (MHD42) [Micah, Maddie, Alex]
Outside Work / Discussion
June 2
Present: Micah, Maddie
Lab Work
- Prepared MHD42 competent cells [Micah, Maddie]
- Checked on transformed MHD42 cells with Uracil DNA Glycosylate 2 [Micah, Maddie]
- Transformation was a success with 100 colonies
- Plate was stored in 4 deg. Celsius for future use
- Prepared TSS Buffer [Micah, Maddie]
Outside Work / Discussion
June 3
No Work!
June 4
No Work!
June 5
Present: Micah, Maddie, Alex, Mark, Collin
Lab Work
- Prepared LB + Chloramphenicol (CM) plates [Micah, Maddie, Mark, Collin]
- Prepared Ampicillin solution to be used for plates in the future [Micah, Maddie, Mark, Collin]
- Made more LB Agar [Alex]
Outside Work / Discussion
- Discussed shifting focus from Dsup gene to a variety of UV radiation resistance gene [Micah, Maddie, Mark, Collin, Alex]
- Found more genes to compare to Dsup (i.e. phrA in tardigrades and cyanobacteria) [Micah, Maddie, Mark, Collin, Alex]
- Met with Dr. Brennan to discuss updates [Micah, Maddie, Mark, Collin, Alex]
June 6
Present: Micah, Maddie, Alex, Mark, Collin
Lab Work
- Prepared DH5α competent cells [Micah, Maddie, Mark, Collin, Alex]
- Tested MHD42 cells for competency [Micah, Maddie, Mark, Collin, Alex]
- Tested DH5α cells for competency [Micah, Maddie, Mark, Collin, Alex]
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 [Micah, Maddie, Mark, Collin, Alex]
- 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) [Micah, Maddie, Mark, Collin, Alex]
June 7
Present: Micah, Maddie, Mark, Collin, Alex
Lab Work
- Checked competency test for MHD42 cells [Micah, Maddie, Mark, Collin, Alex]
- Test was successful!
- Checked competency test for DH5α [Micah, Maddie, Mark, Collin, Alex]
- Test showed that the cells were competent but had a low transformation efficiency
- Prepared LB + Ampicillin (Amp) plates [Micah, Mark, Collin]
- Resuspeded parts BBa_B0034, BBa_R0010, and BBa_K1499200 from current and past distribution kits [Maddie, Alex]
- Transformed part BBa_B0034 (with plasmid psB1A2) into MHD42 competent cells [Maddie, Alex]
- Transformed part BBa_R0010 (with plasmid psB1C3) into MHD42 competent cells [Maddie, Alex]
- Transformed part BBa_K1499200 (with plasmid psB1C3) into MHD42 competent cells [Maddie, Alex]
- Prepared overnight culture of DH5α competent cells [Mark, Collin]
Outside Work / Discussion
- Discussed modelling ideas [Micah, Maddie, Mark, Collin]
- Learned how to design primers from Eugene [Micah, Maddie, Mark, Collin, Alex]
June 8
Present: Alex, Maddie, Micah, Collin, Mark, Zoe
Lab Work
- Checked the results of the transformations of BBa_B0034, BBa_R0010, and BBa_K1499200 into MHD42 cells [Alex, Micah, Zoe]
- Made 900 mL of LB media [Collin, Mark, Zoe]
- Prepared DH5α competent cells [Micah, Zoe, Collin, Mark]
- Tested competency of the prepared DH5 [Maddie, Zoe, Mark]
- Prepared three separate overnight cultures of MHD42 cells with parts BBa_B0034, BBa_R0010, and BBa_K1499200 [Alex, Zoe]
Outside Work / Discussion
- Attended Energy, Environmental, and Chemical Engineering (EECE) summer interns orientation [Collin, Mark, Maddie, Zoe, Alex, Micah]
- Discussed each member's designated role and plans for the future [Collin, Mark, Maddie, Zoe, Alex, Micah]
- Made a team calendar [Collin, Mark, Maddie, Zoe, Alex, Micah]
June 9
Present: Maddie, Zoe, Micah, Mark, Collin, Alex
Lab Work
- Checked results for competency of DH5α cells [Alex, Mark, Micah, Maddie, Zoe, Collin}
- Test was successful! The cells had a moderate transformation efficiency.
- Began the Interlab study by transforming the parts into DH5α cells [Maddie, Zoe, Mark]
- Parts: Postive control (BBa_I20270), Negative control (BBa_R0040), Test device 1 (BBa_J364000), Test device 2 (BBa_J364001), Test device 3 (BBa_J364002), Test device 4 (BBa_J364003), Test device 5 (BBa_J364004), Test device 6 (BBa_J364005)
- Made 400 mL of LB Agar [Zoe]
- Made 11 Chloramphenicol (CM) + LB plates [Collin, Mark, Alex]
Outside Work / Discussion
- Researched modelling ideas [Micah]
June 10
Present: Zoe
Lab Work
- Checked on the transformations for the interlab study [Zoe]
- Colonies existed on all plates. Only a few colonies appeared on the plate with Test Device 1 (BBa_J364000)
- Top (from left to right): Postive control (BBa_I20270), Negative control (BBa_R0040), Test device 1 (BBa_J364000), Test device 2 (BBa_J364001) Bottom (from left to right): Test device 3 (BBa_J364002), Test device 4 (BBa_J364003), Test device 5 (BBa_J364004), Test device 6 (BBa_J364005)
Outside Work / Discussion
June 11
Present: Alex
Lab Work
- Prepared inoculated overnight cultures of the DH5α cells with BBa_B0034, BBa_R0010, and BBa_K1499200 in chloramphenicol [Alex]
Outside Work / Discussion
June 12
Present: Micah, Zoe, Mark, Collin, Maddie, Alex
Lab Work
- Transformed plasmid with GFP into MHD42 competent cells [Zoe, Micah]
- Miniprepped the 3 separate cultures of MHD42 cells that contained BBa_R0010, BBa_B0034, and BBa_K1499200 [Alex, Collin]
- Performed the Calibration Protocol for the Interlab study by measuring the absorbances of distilled water and Ludox HS-40 [Maddie, Mark]
- Measured the fluorescence of fluorescein serial dilutions for the Interlab study [Mark, Maddie]
Outside Work / Discussion
- Filled out Forms 1 and 2 for the Interlab study [Maddie, Mark]
- Studied BioBrick assembly methods [Alex, Micah]
- Met with Dr. Brennan and discussed our project thus far with her [Micah, Collin, Alex, Maddie, Mark, Zoe]
June 13
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