Team:CSU Fort Collins/Experiments
Experiments and daily lab notebook
05-16: Transformed pHF10 and pLC71 into TS559
5-311: Colonies Alive
05-24: Followed spot plate protocol with pHF10
05-29: Transferred colonies to 40 individual 5uL flasks using lab stock ASW media and ASW Additives, Sulfur was not added.
5-30: Sulfur Added to cultures to 20 of those cultures
5-31: No growth on pHF10(1)
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.
6-02: Ran a PCR and gel on the gprepped DNA samples from 6-01.
6-03:Serum bottles checked at 11:10am
6-04: Serum bottles checked at 1:30
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).
6-06: Some oxidized sulfur, but not enough in 6-05 strain. Picked colonies from old PLC71 streak plate into 10 5 mL bottles
6-07: No growth on pLC71
6-08: Started a 100 mL culture of TS559 for transformation. Made minimal ASW plates.
6-09: Performed transformation protocol with pLC71 plasmid
6-14: Made rich ASW plates. Spotted 10 colonies per plate onto the rich ASW media
6-17: Checked plates, but needed more time
6-19: Took 5 5mL serum bottles per plate, and placed in incubator at 11:00 am.
6-20: At 2:30, 9 cultures had grown.
6-21: Performed a gPREP of 3pLC71 cultures and 1 pHF10 culture. Performed pCR protocol.
6-23: Made a gel and and ran the gPrepped cultures
6-26: Started 2 100 mL bottles from pLC71 cultures, 2 bottles from pHF10(1), and two bottles of TS559.
6-27: Removed cultures at 7:15 am. Performed sonication protocol. Stored protein lysate in fridge.
7-6: Performed a Bradford Assay on protein lysate
7-12: Performed a western blot analysis according to protocol. Blott was not clear, and no reasonable results obtained.
7-17: Made ASW media.
7-19 No grotwth on current cultures, started 3 each serum bottles of pLC71 and 3 each of 4 pHF10 strains
7-25: Performed a Gprep on pLC71 and pHF10 samples
8-8: Inoculated 3, 100 mL ASW bottles with 1 mL cultures each (1 TS559, 1 pLC71, 1 pHF10)
8-9: No growth. Used a bottle of laboratory stock media and inoculated with 9 pHF10 strains, 10 pLC71 strains.
8-10: 11:00 am No growth
8-16: Most cultures showed signs of growth.
9-14:Inoculated 4, 100 mL bottles of media ( 2 pLC71, 2 pHF10)
9-16: Made new ASW media.
8-17: Inoculated 4 100 mL bottles of ASW media with 1 TS559, 2 pHF10, and 1 pLC71.
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)
9-26: No growth in any culture.
