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LAB TIME

05-16: Transformed pHF10 and pLC71 into TS559

Courtney

5-311: Colonies Alive Courtney 05-24: Followed spot plate protocol with pHF10 Courtney, Kayla, Frankie, Cooper, Neil 05-29: Transferred colonies to 40 individual 5uL flasks using lab stock ASW media and ASW Additives, Sulfur was not added. Carlos 5-30: Sulfur Added to cultures to 20 of those cultures Carlos 5-31: No growth on pHF10(1) Frankie, Kayla 6-01: Four pHF10(1) cultures survived. Performed a gPrep on these cultures. Made new 25mM lovastatin stock. Used 6-24 pHF10(2) spot plates to make 20 5uL serum bottle cultures via ASW Additives Protocol. Frankie, Neil, Cooper, Kayla 6-02: Ran a PCR and gel on the gprepped DNA samples from 6-01. Kayla, Cooper, Frankie, Melissa 6-03:Serum bottles checked at 11:10am Melissa 6-04: Serum bottles checked at 1:30 Carlos 6-05: Primers for 6-02 gel were wrong, redid gel with 6-01 DNA. Surviving pLC71(2) colonies removed from incubator. Made ASW-yt media. Added additives to 2 100 mL bottles of ASW-yt and inoculated from bottle 2C from pLC71(1). Robyn, Frankie Kayla, Melissa, Cooper 6-06: Some oxidized sulfur, but not enough in 6-05 strain. Picked colonies from old PLC71 streak plate into 10 5 mL bottles Kayla, Carlos, Cooper, Neil 6-07: No growth on pLC71 Kayla, Neil, Carlos, Melissa 6-08: Started a 100 mL culture of TS559 for transformation. Made minimal ASW plates. Frankie, Kayla 6-09: Performed transformation protocol with pLC71 plasmid Frankie, Kayla 6-14: Made rich ASW plates. Spotted 10 colonies per plate onto the rich ASW media Kayla, Courtney 6-17: Checked plates, but needed more time Qianhui, Carlos 6-19: Took 5 5mL serum bottles per plate, and placed in incubator at 11:00 am. Kayla 6-20: At 2:30, 9 cultures had grown. Kayla 6-21: Performed a gPREP of 3pLC71 cultures and 1 pHF10 culture. Performed pCR protocol. Cooper, Carlos, Jake, Robyn, Qianhui, Kayla 6-23: Made a gel and and ran the gPrepped cultures Kayla, Jack 6-26: Started 2 100 mL bottles from pLC71 cultures, 2 bottles from pHF10(1), and two bottles of TS559. Courtney, Skylar 6-27: Removed cultures at 7:15 am. Performed sonication protocol. Stored protein lysate in fridge. Kayla 7-6: Performed a Bradford Assay on protein lysate Neil, Kayla 7-12: Performed a western blot analysis according to protocol. Blott was not clear, and no reasonable results obtained. Melissa, Carlos,Neil, Frankie, Robyn, Kayla 7-17: Made ASW media. Melissa, Kayla 7-19 No grotwth on current cultures, started 3 each serum bottles of pLC71 and 3 each of 4 pHF10 strains Kayla, Melissa, Frankie 7-25: Performed a Gprep on pLC71 and pHF10 samples Kayla, Madeline 8-8: Inoculated 3, 100 mL ASW bottles with 1 mL cultures each (1 TS559, 1 pLC71, 1 pHF10) Neil, Carlos 8-9: No growth. Used a bottle of laboratory stock media and inoculated with 9 pHF10 strains, 10 pLC71 strains. Kayla 8-10: 11:00 am No growth Kayla 8-16: Most cultures showed signs of growth. Kayla 9-14:Inoculated 4, 100 mL bottles of media ( 2 pLC71, 2 pHF10) Kayla 9-16: Made new ASW media. Neil, Kayla 8-17: Inoculated 4 100 mL bottles of ASW media with 1 TS559, 2 pHF10, and 1 pLC71. Kayla 8-19: No growth on cultures. Left in incubator. Made new, smaller cultures using 1 bottle ASW YT media according to protocol (3 bottles pLC71, 12 bottles pHF10) Kayla 9-26: No growth in any culture. Kayla