Difference between revisions of "Part:BBa K824012"

 
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We digested the promoter responsive to Lead (BBa_I721001) with EcoRI and SpeI and digested the GFP plasmid (BBa_I13401) with XbaI and PstI. The promoter and reporter were Ampicillin resistant. These digested fragments were mixed and ligated to the provided, linearized pSB1C3 plasmid. The ligation mix was grown under Chloramphenicol selection. The resulting colonies were tested for responsive GFP production following the addition of Lead Nitrate.
 
We digested the promoter responsive to Lead (BBa_I721001) with EcoRI and SpeI and digested the GFP plasmid (BBa_I13401) with XbaI and PstI. The promoter and reporter were Ampicillin resistant. These digested fragments were mixed and ligated to the provided, linearized pSB1C3 plasmid. The ligation mix was grown under Chloramphenicol selection. The resulting colonies were tested for responsive GFP production following the addition of Lead Nitrate.
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OD of samples were measured at 600 nm along with fluorescence for each sample well. Increasing amounts of lead (II) acetate trihydrate were administered. Relative fluorescence of cells without lead was 2549.89.
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<img src="https://2019.igem.org/wiki/images/7/7b/T--british_columbia--registry-1.png">
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 03:55, 22 October 2019


Lead Promoter with GFP Reporter (Lead Detector)

We digested the promoter responsive to Lead (BBa_I721001) with EcoRI and SpeI and digested the GFP plasmid (BBa_I13401) with XbaI and PstI. The promoter and reporter were Ampicillin resistant. These digested fragments were mixed and ligated to the provided, linearized pSB1C3 plasmid. The ligation mix was grown under Chloramphenicol selection. The resulting colonies were tested for responsive GFP production following the addition of Lead Nitrate.

OD of samples were measured at 600 nm along with fluorescence for each sample well. Increasing amounts of lead (II) acetate trihydrate were administered. Relative fluorescence of cells without lead was 2549.89.

<img src="T--british_columbia--registry-1.png">


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 744
    Illegal SapI.rc site found at 80