Team:Grenoble-EMSE-LSU/Documentation/Notebook/August

From 2013.igem.org

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                                               <h3>Thursday</h3>
                                               <h3>Thursday</h3>
                                                   <h4> KillerRed Characterization </h4>
                                                   <h4> KillerRed Characterization </h4>
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<p> Tested different loading dyes enabling to follow cell viability by fluorescence microscopy. The ON culture of M15[pRep4-pQE30::KR] was split in two different Erlenmeyers and incubated at 37°C, 300rpm. One of them was kept in the dark while the other was illuminated (I = ) for 2 hours to induce cell death. For both samples, 200 µL were pipetted, centrifuged, and incubated 15 min (37°C, 200 rpm) with 200µL of one of the following loading dyes : 5-100µM Diaminofluorescein, 500-5nM MitoTracker Green FM (M7514, Invitrogen, Carlsbad, NM, USA), XX SYBR Safe DNA gel stain (S33102, Invitrogen). Cells were subsequently washed in 200 µL PBS, mounted on a microscope slide and observed with the epifluorescence microscope available in our lab (FITC filter cube).
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<p> Tested different loading dyes enabling to follow cell viability by fluorescence microscopy. </p>
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                                                    <h5> Materials and Methods </h5>
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<p>
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The ON culture of M15[pRep4-pQE30::KR] was split in two different Erlenmeyers and incubated at 37°C, 300rpm. One of them was kept in the dark while the other was illuminated (I = ) for 2 hours to induce cell death. For both samples, 200 µL were pipetted, centrifuged, and incubated 15 min (37°C, 200 rpm) with 200µL of one of the following loading dyes : 5-100µM Diaminofluorescein, 500-5nM MitoTracker Green FM (M7514, Invitrogen, Carlsbad, NM, USA), XX SYBR Safe DNA gel stain (S33102, Invitrogen). Cells were subsequently washed in 200 µL PBS, mounted on a microscope slide and observed with the epifluorescence microscope available in our lab (FITC filter cube).</p>
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                                                    <h5> Results </h5>
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<p></p>
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<p> Performed a new kinetics on M15[pRep4-pQE30::KR] cells, in M9 medium. </p>
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                                                    <h5> Materials and Methods </h5>
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<p>
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The ON culture of M15[pRep4-pQE30::KR] was split in two different Erlenmeyers and incubated at 37°C, 300rpm. One of them was kept in the dark while the other was illuminated (I = ) for 2 hours to induce cell death. For both samples, 200 µL were pipetted, centrifuged, and incubated 15 min (37°C, 200 rpm) with 200µL of one of the following loading dyes : 5-100µM Diaminofluorescein, 500-5nM MitoTracker Green FM (M7514, Invitrogen, Carlsbad, NM, USA), XX SYBR Safe DNA gel stain (S33102, Invitrogen). Cells were subsequently washed in 200 µL PBS, mounted on a microscope slide and observed with the epifluorescence microscope available in our lab (FITC filter cube).</p>
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                                                    <h5> Results </h5>
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<p></p>
                                               <h3>Friday</h3>
                                               <h3>Friday</h3>
                                                                                      
                                                                                      

Revision as of 14:19, 22 August 2013

Grenoble-EMSE-LSU, iGEM


Grenoble-EMSE-LSU, iGEM

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