Difference between revisions of "Part:BBa M36661:Experience"

Revision as of 20:26, 6 December 2013

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Applications of BBa_M36661

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Stanford Location

Plasmid Name: fur protein, DNA 2.0 Gene #: 135239, Organism: K12 E.coli, Device Type: Sensor, Box Label: BIOE 44 F13, Barcode #'s of Glycerol Stocks: 0133011789, 0113010769, 0133009689, 0133011828, 0133009694, 0133011775(This one used to Restart Stock for Fur Sensor Project)

User Reviews

UNIQ7f28c610e01966bc-partinfo-00000001-QINU ANOVA of BETA GAL Round 1—p=.239 NOT SIGNIFICANT ANOVA of BETA GAL Round 2—p=.389 NOT SIGNIFICANT

Student t-test Beta Gal round 2 -4 vs -7: t= -1.43 sdev= 6.85 degrees of freedom = 14 The probability of this result, assuming the null hypothesis, is 0.18(Not significant)

-4 vs no iron: t= -1.80 sdev= 9.15 degrees of freedom = 12 The probability of this result, assuming the null hypothesis, is 0.097( not significant)

-7 vs no iron: t=-0.823 sdev= 9.03 degrees of freedom = 12 The probability of this result, assuming the null hypothesis, is 0.43(Not significant)

Figure #1 (above):Relationship of the Beta-Gal production from the second test. Each category contained n=4 duplicates. Values were averaged and standard deviations were calculated to develop the above graph. Analysis with One-Way ANOVA (p=.389) and Student t-tests between groups showed no statistical significance.

Figure #2 (above):Relationship of the Beta-Gal production from the first round of experimentation. Each category contained n=2 duplicates. Values were averaged and standard deviations were calculated to develop the above graph. Analysis with One-Way ANOVA (p=.2399) showed no statistical significance. (NOTE: There were too few duplicates per category to perform Student t-tests between groups.)

Our second round of experimentation did not yield a significant difference between groups, suggesting that our construct is not functional.

The results from this first round showed a promising difference between the iron concentration of 10^-4 M and no iron, as we expected. This led us to develop a second round of experimentation for look for statistical differences with more samples. Since there was not a significant difference between the groups, we also steered away from looking for a gradient and spent our efforts testing for functionality in the second round of experimentation.

Figure #3: (above) The results from Gel electrophoresis demonstrated successful transcription (shown by the bright bands). The results also suggest the presence of our DNA, however the faint bands and our inability to identify the copy number of our construct (due to an unexpected mutation) make these results unclear.

Figure #4: (above) Very faint fluorescence some potential expression (I’m not sure if you want to include this one or not…I think it might be better to show the two below showing that our plasmid was indeed taken up)

Figure #5: (above) Amp plate—plasmid was taken up.

Kan plate—no growth as expected UNIQ7f28c610e01966bc-partinfo-00000002-QINU

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 [[Image:pcr1.png]]
-Figure #3: (above) Gel shows bright bands in our RT PCR demonstrating successful transcription and faint bands in the DNA length we were looking for, suggesting the presence of fur DNA.
+Figure #3: (above) The results from Gel electrophoresis demonstrated successful transcription (shown by the bright bands). The results also suggest the presence of our DNA, however the faint bands and our inability to identify the copy number of our construct (due to an unexpected mutation) make these results unclear.
 [[Image:plate1.png]]