Team:Nevada/project/OMpermeabilization

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(Results)
 
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::Sequencing data revealed that we had deleted the appropriate region  
::Sequencing data revealed that we had deleted the appropriate region  
::This construct was transformed into BL21 cells and induced with 100mM IPTG
::This construct was transformed into BL21 cells and induced with 100mM IPTG
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Results: This design resulted in the deletion of 372 base pairs yielding a 2070 base pair construct of Plac-INPN-YFP as evidenced by gel electrophoresis (figure 2). Fluorescence observed under fluorescent microscopy at equivalence to Edinburgh’s construct.  
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Results: This design resulted in the deletion of 372 base pairs yielding a 2070 base pair construct of Plac-INPN-YFP as evidenced by gel electrophoresis (figure 10). Fluorescence observed under fluorescent microscopy at equivalence to Edinburgh’s construct.  
[[File:nvresults9.jpg|thumb|center]]   
[[File:nvresults9.jpg|thumb|center]]   
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Figure 1: Plca-INPN-YFP Plasmid map
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Figure 9: Plca-INPN-YFP Plasmid map
[[File:nvresults10.jpg|thumb|center]]   
[[File:nvresults10.jpg|thumb|center]]   
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Figure 2: 1% agarose of PstI and EcorI digested Plac-INPN-YFP and Plac-INPNC-YFP: Lane 1 is a 1kb marker. Lanes 2 and 3 are PstI and EcoRI digested pSB1C3 with expected construct at 2070 bp.  pSB1C3 is also 2070 bp therefore the correct construct will show as one band with the backbone. Lane 4 is PstI and EcoRI digested pSB1C3 with the original Plac-INPNC-YFP construct. Lanes 5 and 6 are undigested Plac-INPN-YFP and Plac-INPNC-YFP respectfully.  
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Figure 10: 1% agarose of PstI and EcorI digested Plac-INPN-YFP and Plac-INPNC-YFP: Lane 1 is a 1kb marker. Lanes 2 and 3 are PstI and EcoRI digested pSB1C3 with expected construct at 2070 bp.  pSB1C3 is also 2070 bp therefore the correct construct will show as one band with the backbone. Lane 4 is PstI and EcoRI digested pSB1C3 with the original Plac-INPNC-YFP construct. Lanes 5 and 6 are undigested Plac-INPN-YFP and Plac-INPNC-YFP respectfully.
==Design 2==  
==Design 2==  
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==Results==  
==Results==  
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This design resulted in the deletion of 622 base pairs yielding a 1448 base pair construct of INP-N-YFP as evidenced by gel electrophoresis (figure 4). No fluorescence was detected in this design.  
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This design resulted in the deletion of 622 base pairs yielding a 1448 base pair construct of INP-N-YFP as evidenced by gel electrophoresis (figure 11). No fluorescence was detected in this design.  
[[File:nvresults10.jpg|thumb|center]]  
[[File:nvresults10.jpg|thumb|center]]  
Figure 11: 1% agarose of PCR amplified INPN-YFP in lane 2 against a 1 kb marker in lane 1. Expected band size of 1448 observed.
Figure 11: 1% agarose of PCR amplified INPN-YFP in lane 2 against a 1 kb marker in lane 1. Expected band size of 1448 observed.

Latest revision as of 04:09, 28 September 2013