MIT7_13f08_lab11_Protocol_Motility

MIT7_13f08_lab11_Protocol_Motility - MIT OpenCourseWare...

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MIT OpenCourseWare http://ocw.mit.edu 7.13 Experimental Microbial Genetics Fall 2008 For information about citing these materials or our Terms of Use, visit: http://ocw.mit.edu/terms .
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7.13 Fall 2008 Page | 1 MOTILITY ASSAYS Twitching Concept - P. aeruginosa cells can adhere to and move across solid surfaces by extending and retracting type IV pili, a behavior known as ‘twitching’. Plates - LB + 1% bacto-agar o pour as shallow a layer of LB/agar as possible; this will aid in visualizing the twitching zone Assay - grow liquid culture o/n - aspirate 2ul culture and insert pipette tip through the gel pad to the plastic/gel interface - dispense 2ul culture into the interface o move pipette tip very slightly side to side until liquid volume is completely dispensed - incubate plates for 24h at 37C Result - strains with functional pili will form a large, haze of growth surrounding a smaller, thicker colony o plates can be held up to light to aid in visualizing the twitching zone left : PA14 pilB::MAR2XT7 (DKN# 599) right : PA14 (DKN# 263)
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7.13 Fall 2008
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MIT7_13f08_lab11_Protocol_Motility - MIT OpenCourseWare...

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