Difference between revisions of "Part:BBa J100308:Experience"

 
 
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===Applications of BBa_J100308===
 
===Applications of BBa_J100308===
  
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We tested E. coli containing either J100306 or its variant, J100308, and measured the induction caused by the addition of 200 ng/mL anhydrous tetracycline (aTc) to determine the incubation time needed for maximum induction. We found that aTc is most effective after fourteen to sixteen hours of growth, after which the induction is lost.
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We performed a dose-response test on E. coli containing part J100306 to determine the ideal concentration of aTc for induction of RFP. We found that 400 ng/mL aTc gives the best compromise of fold induction and allowing for sufficient cell growth. Since J100308 and J100306 responded similarly in the time-response test, we expect that 400 ng/mL aTc would also be ideal for J100308 and other repClone constructs built from the variant J100307. Induction above 400 ng/mL looks higher because fewer cells grew and therefore the denominator was smaller when calculating the ratio. 
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Results of Dose Response Assay:
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[[File:J100306DoseResponseTable.png]]
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[[File:J100306DoseResponseInductionRatios.png]]
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Results of Time Assay:
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[[File:repCloneTimeAssay.png]]
 
===User Reviews===
 
===User Reviews===
 
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Latest revision as of 18:57, 17 October 2016


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Applications of BBa_J100308

We tested E. coli containing either J100306 or its variant, J100308, and measured the induction caused by the addition of 200 ng/mL anhydrous tetracycline (aTc) to determine the incubation time needed for maximum induction. We found that aTc is most effective after fourteen to sixteen hours of growth, after which the induction is lost.

We performed a dose-response test on E. coli containing part J100306 to determine the ideal concentration of aTc for induction of RFP. We found that 400 ng/mL aTc gives the best compromise of fold induction and allowing for sufficient cell growth. Since J100308 and J100306 responded similarly in the time-response test, we expect that 400 ng/mL aTc would also be ideal for J100308 and other repClone constructs built from the variant J100307. Induction above 400 ng/mL looks higher because fewer cells grew and therefore the denominator was smaller when calculating the ratio.

Results of Dose Response Assay:


J100306DoseResponseTable.png


J100306DoseResponseInductionRatios.png


Results of Time Assay:


RepCloneTimeAssay.png

User Reviews

UNIQ0470d56c2c5ee4dd-partinfo-00000000-QINU UNIQ0470d56c2c5ee4dd-partinfo-00000001-QINU