Difference between revisions of "Part:BBa K412000:Experience"

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<partinfo>BBa_J23109 AddReview 5</partinfo>
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Results of nitrate induction are presented on the left. (A) It is obvious that nitrate can induce GFP expression. Additionally, Pyear (nitrate-sensitive promoter) respond ultra-sensitively when nitrate concentration ranges from approximately 0-2 ug/L. And after concentration of 3.5ug/L,GFP expression maintains at nearly 2800 RFU which showcase its maximal initiation rate of promoter. (B)Top10 brand control is presented. We can clearly see the differences between (A) and (B).
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Latest revision as of 14:46, 26 September 2012

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_K412000

Ver2.jpg

  • All images were taken with Olympus IX81 automated inverted microscope specially equipped for live cell imaging. All experiments were performed with DH5α E.coli cells. The filter sets we used are: 545/30x (excitation) and 620/60m (emission) filters for DsRed, 470/40x (excitation) and 525/50m (emission) for GFP. Data collection and processing was performed by the SlideBook software.
  • The data was taken using cells that were grown overnight in 2mL of LB broth that had 50ug/mL of the appropriate antibiotic at 37C and 220rpm. This was used in a 1:50 dilution. Each sample had 40uL from the broth grown overnight and 2mL of the LB broth with the appropriate antibiotic. These were allowed to grow until a predetermined OD. Then the contaminants were added and the cells were allowed to grow for another 2 hours, before the images were taken.
  • The images shown are the images used to get the data. From left to right the image is the negative control to a 10mM concentration of the respective contaminants. The graph shows the average of all the maximum intensities of the cells in the image vs the concentration of nitrate/nitrite added to the sample. The data shows that PyeaR+GFP responds better to nitrates than nitrites. It also shows that the cell is more sensitive to aluminum nitrate nonahydrate, and therefore starts dying at a lower concentration.

Tileview.jpg

The images above are timelapse images. The images were taken every ten minutes after the 10mM potassium nitrate was added to the sample.

User Reviews

UNIQ46ae65e3e5bd5f8f-partinfo-00000000-QINU UNIQ46ae65e3e5bd5f8f-partinfo-00000001-QINU

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iGEM12_OUC-China

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800px-N22.PNG

Results of nitrate induction are presented on the left. (A) It is obvious that nitrate can induce GFP expression. Additionally, Pyear (nitrate-sensitive promoter) respond ultra-sensitively when nitrate concentration ranges from approximately 0-2 ug/L. And after concentration of 3.5ug/L,GFP expression maintains at nearly 2800 RFU which showcase its maximal initiation rate of promoter. (B)Top10 brand control is presented. We can clearly see the differences between (A) and (B).