Chaston:Notebook/chaston/2010/03/02

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2010/03/02 Data UV NilB • Verify UV susceptibility • qRT-PCR of XaxAB, also maybe xhlA and the other hemolysins – talk to XJ 5 - NilB Homologs • Clone new NilB Homologs • Analyze Context of NilB Homologs • Sequence some strains NilA FLAG • Constitutive NilB Expression • Creating NilA-FLAG strains o Boil prep on nilAFLAG-2, -25, -50, -75, -90, and 50-75 in 1430 o Do Tn7-detect o Run gel o Streak strains (put plates out for streaking) o Prep minimal medium so that I can subculture the cells and have them ready to test for western 4 - NilB Deletion • qPCR o analyze the data CM1Z • Assess colonization of FLAG strains o Type up and process data for low-detection colonization assays Archna Science • Assess colonization of FLAG strains o Do Tn7 detect on 1406-292 and -315 o Run gel o Streak strains (put plates out for streaking) Nematode Colonization • Compete RFP vs. GFP WT o Move samples to water traps • Do bacteria colonize eggs o Check samples, record observations • Chase experiments o Check samples, record observations Western • Run westerns o Figure out protein concentrations to run o Make 7.5% SDS-PAGE gels tonight o • 6- Prep western samples • 6 - Prep Para-nilB samples • 6 - Make minimal medium (1.11X) • Streak cultures • qPCR • 7 - Try to figure out how to do something with glucose-6-phosphate. Look on the internet for recipes doing that. • Count, by epifluorescent microscopy, the GFP colonization of high numbers of nematodes of my competition assays • Put together pictures to tell a story • PCR pEVS107-R1 and SR1-Before1 in 281 11F; also check if it is strep resistant