Difference between revisions of "Part:BBa J33203:Experience"

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===Applications of BBa_J33203===
 
===Applications of BBa_J33203===
  
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In 2010, Team Debrecen-Hungary has decided to give a hand with testing the BBa_J33203 part. As the South-East of Hungary is highly affected by the arsenic problem (there are wells in which the level of arsenic is at least 15-fold higher than the WHO limit of 10 ug/l), we had the possibility to use real-world samples for testing.
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Team Debrecen-Hungary has decided to give a hand with testing the BBa_J33203 part. As the South-East of Hungary is highly affected by the arsenic problem (there are wells in which the level of arsenic exceeds the allowed limit), we had the possibility to use real-world samples for testing.
  
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We collected samples from two wells which were known to contain elevated arsenic concentration. We used DH5 alpha E. coli cell line with lacZDM15 mutation as host cells, than we prepared a medium consisting of ''peptone, yeast extract, NaCl, K2HPO4, NaHCO3, bromthymol blue, and autoclaved water samples of different dilutions''. We spiked these media with the transformed cells, and after 24 hours of incubation at 37°C we received that the part works also on real world samples, not only on samples made of water and sodium-arsenate. The colour change of Bromothymol Blue seemed to be directly proportional to the concentration of arsenic.
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We collected samples from two wells which were known to contain elevated arsenic concentration (Szegfű and Madách street, Békéscsaba, Hungary). We used DH5 alpha E. coli cell line with lacZDM15 mutation as host cells, than we prepared a medium consisting of ''peptone, yeast extract, NaCl, K2HPO4, NaHCO3, bromthymol blue, and autoclaved water samples of different dilutions''. We spiked these media with the transformed cells, and after 24 hours of incubation at 37°C we received that the part works also on real world samples, not only on samples made of water and sodium-arsenate. The colour change of Bromothymol Blue seemed to be directly proportional to the concentration of arsenic. The reference dilutions made from Na-arsenate and MilliQ water shows that the concentration of arsenic in our collected samples is at least 15x higher than the WHO recommended limit.
  
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[[Image:Arsenic_before incubation]]
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<div style="overflow: auto; width: 100%;">
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<div style="float: left;">
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[[Image:Refdilarsenic.jpg|thumb|430x310px|Reference dilutions from Na-arsenate/after 24 hours of incubation.]]
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</div>
  
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<div style="float: right;">
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[[Image:Arsenic Szegfu street after 24 hours of incubation wikire.jpg|thumb|430x310px|Sample from Szegfű street/after 24 hours of incubation.]]
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</div>
  
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[[Image:arsenic_Madách street_after 24 hours of incubation]]
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<div style="float: left;">
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[[Image:Arsenic_Madach street after 24 hours of incubation wikire.jpg|thumb|430x310px|Sample from Madách street/after 24 hours of incubation.]]
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</div>
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<div style="clear: both;"></div>
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<div>
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These samples were used for the publication of the Arsenic Biosensor by the members of the Edinburgh Team.
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Reference:
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''Anal Bioanal Chem. 2011 May;400(4):1031-9. Epub 2011 Mar 27.''
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'''''A pH-based biosensor for detection of arsenic in drinking water.'''''
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''de Mora K, Joshi N, Balint BL, Ward FB, Elfick A, French CE.'',
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''School of Engineering, University of Edinburgh, UK.''
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For more info about source of the samples or experiments, please contact Lilla Ozgyin (iGEM Team Debrecen_Hungary 2011)
  
 
===User Reviews===
 
===User Reviews===

Latest revision as of 16:22, 21 September 2011

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_J33203

Team Debrecen-Hungary has decided to give a hand with testing the BBa_J33203 part. As the South-East of Hungary is highly affected by the arsenic problem (there are wells in which the level of arsenic exceeds the allowed limit), we had the possibility to use real-world samples for testing.

We collected samples from two wells which were known to contain elevated arsenic concentration (Szegfű and Madách street, Békéscsaba, Hungary). We used DH5 alpha E. coli cell line with lacZDM15 mutation as host cells, than we prepared a medium consisting of peptone, yeast extract, NaCl, K2HPO4, NaHCO3, bromthymol blue, and autoclaved water samples of different dilutions. We spiked these media with the transformed cells, and after 24 hours of incubation at 37°C we received that the part works also on real world samples, not only on samples made of water and sodium-arsenate. The colour change of Bromothymol Blue seemed to be directly proportional to the concentration of arsenic. The reference dilutions made from Na-arsenate and MilliQ water shows that the concentration of arsenic in our collected samples is at least 15x higher than the WHO recommended limit.

Reference dilutions from Na-arsenate/after 24 hours of incubation.
Sample from Szegfű street/after 24 hours of incubation.
Sample from Madách street/after 24 hours of incubation.

These samples were used for the publication of the Arsenic Biosensor by the members of the Edinburgh Team.

Reference:

Anal Bioanal Chem. 2011 May;400(4):1031-9. Epub 2011 Mar 27.

A pH-based biosensor for detection of arsenic in drinking water.

de Mora K, Joshi N, Balint BL, Ward FB, Elfick A, French CE., School of Engineering, University of Edinburgh, UK.


For more info about source of the samples or experiments, please contact Lilla Ozgyin (iGEM Team Debrecen_Hungary 2011)

User Reviews

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