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


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