Difference between revisions of "Part:BBa K216016:Experience"
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| − | '''Initial tests: Edinburgh iGEM 2009:''' This part was tested by growing ''E. coli'' JM109/pSB1A2-BBa_K216016 overnight at 37 C with shaking in LB with 80 mg/l ampicillin and varying concentrations of nitrate and nitrite (0, 5, 10 mM sodium nitrite, and 10, 20, 40 mM sodium nitrate). | + | '''Initial tests: Edinburgh iGEM 2009:''' This part was tested by growing ''E. coli'' JM109/pSB1A2-BBa_K216016 overnight at 37 C with shaking in LB with 80 mg/l ampicillin and varying concentrations of nitrate and nitrite (0, 5, 10 mM sodium nitrite, and 10, 20, 40 mM sodium nitrate). The following day, 0.05% (ie 0.5 microlitres/ml) n-decanal was added, and the vials were observed in a dark room. The control, with no nitrate or nitrate, was least bright, possibly non-luminescent. The brightest was the vial with 20 mM nitrate, followed by that with 10 mM nitrate. The others, with 40 mM nitrate or 5 or 10 mM nitrite, were of intermediate brightness. Unfortunately, we don't currently have a luminometer to make a quantitive measurement of luminescence, but in qualitative terms, this part seems to work fine. |
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| + | [[Image:XL_LuxAB.jpg]] | ||
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Latest revision as of 15:19, 21 October 2009
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_K216016
This part is designed to produce a luminescent response in the presence of nitrate or nitrite anions. The bacterial luciferase reaction also requires oxygen and a long chain aldehyde. This can be biosynthesised by the luxCDE gene products (we are preparing a BioBrick with these) or can be supplied in the form of 0.05 - 0.1% v/v n-decanal. Luminescence is blue, around 495 nm (see notes on part BBa_K216008 for more details).
User Reviews
Initial tests: Edinburgh iGEM 2009: This part was tested by growing E. coli JM109/pSB1A2-BBa_K216016 overnight at 37 C with shaking in LB with 80 mg/l ampicillin and varying concentrations of nitrate and nitrite (0, 5, 10 mM sodium nitrite, and 10, 20, 40 mM sodium nitrate). The following day, 0.05% (ie 0.5 microlitres/ml) n-decanal was added, and the vials were observed in a dark room. The control, with no nitrate or nitrate, was least bright, possibly non-luminescent. The brightest was the vial with 20 mM nitrate, followed by that with 10 mM nitrate. The others, with 40 mM nitrate or 5 or 10 mM nitrite, were of intermediate brightness. Unfortunately, we don't currently have a luminometer to make a quantitive measurement of luminescence, but in qualitative terms, this part seems to work fine.
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