Team:Slovenia/Notebook

From 2012.igem.org

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<h2>Induction systems</h2>
<h2>Induction systems</h2>
The induction systems are described <a href="https://2012.igem.org/Team:Slovenia/Parts">here</a>.  
The induction systems are described <a href="https://2012.igem.org/Team:Slovenia/Parts">here</a>.  
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<p>The induction systems are described here. </p>
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<li>2 hours post transfection change media and stimulate the cells by adding dilutions of appropriate inductors to the medium in a 1:10 (v:v). </li>
<li>2 hours post transfection change media and stimulate the cells by adding dilutions of appropriate inductors to the medium in a 1:10 (v:v). </li>
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<table class="normal" style="font-size:90%; width:90%; text-align:center;">
<table class="normal" style="font-size:90%; width:90%; text-align:center;">
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<h2>Effectors </h2>
<h2>Effectors </h2>
<h3>Biological assay-anakinra </h3>
<h3>Biological assay-anakinra </h3>
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<p>For spatial and temporal imaging of samples a Leica TCS SP5 laser scanning microscope mounted on a Leica DMI 6000 CS inverted microscope (Leica Microsystems, Germany) with a 10× and 20× dry objective and an HCX plan apo 63× oil (NA 1.4) oil immersion objective was used. For image analysis we used ImageJ (Image Processing and Analysis in Java) software (http://rsbweb.nih.gov/ij/) measuring the mean grey values of each cell containing the promoter of interest. </p>
<p>For spatial and temporal imaging of samples a Leica TCS SP5 laser scanning microscope mounted on a Leica DMI 6000 CS inverted microscope (Leica Microsystems, Germany) with a 10× and 20× dry objective and an HCX plan apo 63× oil (NA 1.4) oil immersion objective was used. For image analysis we used ImageJ (Image Processing and Analysis in Java) software (http://rsbweb.nih.gov/ij/) measuring the mean grey values of each cell containing the promoter of interest. </p>
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<h3>Microscopy-cell viability with Hoechst and SytoxGreen514 (Safety mechanisms) </h3>
<h3>Microscopy-cell viability with Hoechst and SytoxGreen514 (Safety mechanisms) </h3>
<p><b>Hoechst</b> dye is a membrane permeable dye and stains all cells in a culture. On the other hand a <b>SytoxGreen514</b> dye is a membrane impermeable dye staining only dead cells. Both dyes, blue fluorescent Hoechst and green fluorescent SytoxGreen514, bind to nucleic acids causing emission of fluorescent light. </p>
<p><b>Hoechst</b> dye is a membrane permeable dye and stains all cells in a culture. On the other hand a <b>SytoxGreen514</b> dye is a membrane impermeable dye staining only dead cells. Both dyes, blue fluorescent Hoechst and green fluorescent SytoxGreen514, bind to nucleic acids causing emission of fluorescent light. </p>
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<p><b>Wash buffer</b>:    1x PBS,    0,01% (v/v) Tween 20</p>
<p><b>Wash buffer</b>:    1x PBS,    0,01% (v/v) Tween 20</p>
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<h2 style="color:grey;">References</h2>
 
<h2 style="color:grey;">References</h2>
<h2 style="color:grey;">References</h2>

Revision as of 18:54, 26 September 2012