Difference between revisions of "Team:Tec-Monterrey/InterLab"

m
 
(107 intermediate revisions by 5 users not shown)
Line 3: Line 3:
 
<head>
 
<head>
 
<style>
 
<style>
−
.div-header-bottom {
+
.header-background {
−
  position: absolute;
+
−
}
+
−
#headerBackground {
+
 
   display: flex;
 
   display: flex;
 
   z-index: 1;
 
   z-index: 1;
Line 16: Line 13:
 
   direction: rtl;
 
   direction: rtl;
 
}
 
}
−
.headerBackgroundLeft, .headerBackgroundCenter, .headerBackgroundCenterRight, .headerBackgroundRight {
+
.header-background-left, .header-background-center, .header-background-center-right, .header-background-right {
 
   width: 18%;
 
   width: 18%;
 
}
 
}
Line 24: Line 21:
 
   top: 5%;
 
   top: 5%;
 
}
 
}
−
/******************************************************** TEST SECTION **************************************/
+
 
−
.seccion_responsiva {
+
  .cd-vertical-nav li:first-of-type a::after {
−
  width: 80%;
+
    /* change icon using image sprites */
−
  background: whitesmoke;
+
    background-position: 0px -48px;
−
  margin: 1% auto;
+
    /* fix bug with transitions - set scale(0.01) rather than scale(0) */
−
}
+
    -webkit-transform: translateX(-50%) translateY(4.5em) scale(0.01);
−
.resumen {
+
    -moz-transform: translateX(-50%) translateY(4.5em) scale(0.01);
−
}
+
    -ms-transform: translateX(-50%) translateY(4.5em) scale(0.01);
−
.introduccion {
+
    -o-transform: translateX(-50%) translateY(4.5em) scale(0.01);
−
}
+
    transform: translateX(-50%) translateY(4.5em) scale(0.01);
−
.contenido {
+
  }
−
}
+
  .cd-vertical-nav li:first-of-type a::before {
−
.contenido_con_imagen_derecha {
+
    -webkit-transform: translateX(-50%) translateY(4.5em) scale(0.25);
−
}
+
    -moz-transform: translateX(-50%) translateY(4.5em) scale(0.25);
−
.contenido_con_imagen_izquierda {
+
    -ms-transform: translateX(-50%) translateY(4.5em) scale(0.25);
−
}
+
    -o-transform: translateX(-50%) translateY(4.5em) scale(0.25);
−
.imagen_centrada_reducida {
+
    transform: translateX(-50%) translateY(4.5em) scale(0.25);
−
}
+
  }
−
.imagen_centrada_completa {
+
  .cd-vertical-nav li:nth-of-type(2) a::after {
−
}
+
    background-position: 0px -32px;
−
.conclusion {
+
    -webkit-transform: translateX(-50%) translateY(1.5em) scale(0.01);
−
}
+
    -moz-transform: translateX(-50%) translateY(1.5em) scale(0.01);
−
.referencias {
+
    -ms-transform: translateX(-50%) translateY(1.5em) scale(0.01);
−
}
+
    -o-transform: translateX(-50%) translateY(1.5em) scale(0.01);
 +
    transform: translateX(-50%) translateY(1.5em) scale(0.01);
 +
  }
 +
  .cd-vertical-nav li:nth-of-type(2) a::before {
 +
    -webkit-transform: translateX(-50%) translateY(1.5em) scale(0.25);
 +
    -moz-transform: translateX(-50%) translateY(1.5em) scale(0.25);
 +
    -ms-transform: translateX(-50%) translateY(1.5em) scale(0.25);
 +
    -o-transform: translateX(-50%) translateY(1.5em) scale(0.25);
 +
    transform: translateX(-50%) translateY(1.5em) scale(0.25);
 +
  }
 +
  .cd-vertical-nav li:nth-of-type(3) a::after {
 +
    background-position: 0px -16px;
 +
    -webkit-transform: translateX(-50%) translateY(-1.5em) scale(0.01);
 +
    -moz-transform: translateX(-50%) translateY(-1.5em) scale(0.01);
 +
    -ms-transform: translateX(-50%) translateY(-1.5em) scale(0.01);
 +
    -o-transform: translateX(-50%) translateY(-1.5em) scale(0.01);
 +
    transform: translateX(-50%) translateY(-1.5em) scale(0.01);
 +
  }
 +
  .cd-vertical-nav li:nth-of-type(3) a::before {
 +
    -webkit-transform: translateX(-50%) translateY(-1.5em) scale(0.25);
 +
    -moz-transform: translateX(-50%) translateY(-1.5em) scale(0.25);
 +
    -ms-transform: translateX(-50%) translateY(-1.5em) scale(0.25);
 +
    -o-transform: translateX(-50%) translateY(-1.5em) scale(0.25);
 +
    transform: translateX(-50%) translateY(-1.5em) scale(0.25);
 +
  }
 +
  .cd-vertical-nav li:nth-of-type(4) a::after {
 +
    background-position: 0px 0px;
 +
    -webkit-transform: translateX(-50%) translateY(-4.5em) scale(0.01);
 +
    -moz-transform: translateX(-50%) translateY(-4.5em) scale(0.01);
 +
    -ms-transform: translateX(-50%) translateY(-4.5em) scale(0.01);
 +
    -o-transform: translateX(-50%) translateY(-4.5em) scale(0.01);
 +
    transform: translateX(-50%) translateY(-4.5em) scale(0.01);
 +
  }
 +
  .cd-vertical-nav li:nth-of-type(4) a::before {
 +
    -webkit-transform: translateX(-50%) translateY(-4.5em) scale(0.25);
 +
    -moz-transform: translateX(-50%) translateY(-4.5em) scale(0.25);
 +
    -ms-transform: translateX(-50%) translateY(-4.5em) scale(0.25);
 +
    -o-transform: translateX(-50%) translateY(-4.5em) scale(0.25);
 +
    transform: translateX(-50%) translateY(-4.5em) scale(0.25);
 +
  }
 +
 
 
</style>
 
</style>
 
</head>
 
</head>
 
<body>
 
<body>
 +
<!------------------------------------------------------------------------------------------------------------------------>
 +
<!----------------------------------------------------- sidebar dots navigation ------------------------------------------>
 +
<!------------------------------------------------------------------------------------------------------------------------>
 +
    <script src="https://2018.igem.org/Team:Tec-Monterrey/T--Tec-Monterrey--modernizr?action=raw&ctype=text/javascript"</script>
 +
    <script src="https://2018.igem.org/Team:Tec-Monterrey/T--Tec-Monterrey--main-vertical-nav?action=raw&ctype=text/javascript"></script>
 +
    <!-- Modernizr -->
 +
   
 +
   
 +
    <!------------SideBar dots navigation -------->
 +
    <nav class="cd-vertical-nav">
 +
        <ul class="vertical-nav">
 +
            <li><a href="#page-top"><span class="label">Header<br><br></span></a></li>
 +
            <li><a href="#procedure-interlab"><span class="label">Procedure<br><br></span></a></li>
 +
            <li><a href="#results-interlab"><span class="label">Results<br><br></span></a></li>
 +
            <li><a href="#conclusion-interLab"><span class="label">Conclusion<br><br></span></a></li>
 +
        </ul>
 +
    </nav>
 +
    <!-- .cd-vertical-nav -->
 +
   
 +
    <script src="https://2018.igem.org/Team:Tec-Monterrey/T--Tec-Monterrey--main?action=raw&ctype=text/javascript"></script>
 +
    <!-- Resource jQuery -->
 +
 
   <!-- Header -->
 
   <!-- Header -->
−
   <header class="masthead" id="interLabHeader">
+
   <header class="masthead" id="page-top">
−
       <div class="header-top" style="background: #397471;">
+
       <div class="header-top">
 +
        <video autoplay muted loop class="video-header" type="video/mp4">
 +
          <source src="https://static.igem.org/mediawiki/2018/f/f9/T--Tec-Monterrey--HeaderTeam.mp4">
 +
        </video>
 
       </div>
 
       </div>
 
       <div class="header-bottom">
 
       <div class="header-bottom">
−
         <div id="headerBackground">
+
         <div class="header-background">
−
           <div class="headerBackgroundRight">
+
           <div class="header-background-right" style="margin-left: -10px; margin-right: 1%;">
 
             <img class="trail" src="https://static.igem.org/mediawiki/2018/5/51/T--Tec-Monterrey--Header_Trail_Far_Right.svg">
 
             <img class="trail" src="https://static.igem.org/mediawiki/2018/5/51/T--Tec-Monterrey--Header_Trail_Far_Right.svg">
 
           </div>
 
           </div>
−
           <div class="headerBackgroundCenterRight">
+
           <div class="header-background-center-right" style="margin-right: -5%; margin-left: -10px;">
 
             <img class="trail" src="https://static.igem.org/mediawiki/2018/7/72/T--Tec-Monterrey--Header_Trail_Center_Right.svg">
 
             <img class="trail" src="https://static.igem.org/mediawiki/2018/7/72/T--Tec-Monterrey--Header_Trail_Center_Right.svg">
 
           </div>
 
           </div>
−
           <div class="headerBackgroundCenter">
+
           <div class="header-background-center" style="margin-left: -10px; margin-right: -10px;">
 
             <img class="trail" src="https://static.igem.org/mediawiki/2018/7/7c/T--Tec-Monterrey--Header_Trail_Center_2.svg">
 
             <img class="trail" src="https://static.igem.org/mediawiki/2018/7/7c/T--Tec-Monterrey--Header_Trail_Center_2.svg">
 
           </div>
 
           </div>
−
           <div  class="headerBackgroundLeft">
+
           <div  class="header-background-left" style="margin-right: -10px;">
 
             <img class="trail" src="https://static.igem.org/mediawiki/2018/a/a6/T--Tec-Monterrey--Header_Trail_Left.svg">
 
             <img class="trail" src="https://static.igem.org/mediawiki/2018/a/a6/T--Tec-Monterrey--Header_Trail_Left.svg">
 
           </div>
 
           </div>
Line 72: Line 134:
 
           <div class="div-header-bottom">
 
           <div class="div-header-bottom">
 
           <div class="header-title">
 
           <div class="header-title">
−
               Interlab
+
               InterLab
 
           </div>
 
           </div>
 
           <div class="header-subtitle">
 
           <div class="header-subtitle">
Line 81: Line 143:
 
   </header>
 
   </header>
  
−
   <!-- Test -->
+
   <div class="articulo">
−
  <section id="testSection" class="seccion_responsiva">
+
    <section id="procedure-interlab" class="seccion-responsiva">
−
    <div class="resumen">
+
      <div class="articulo">
−
      Lorem ipsum dolor sit amet, consectetur adipiscing elit. Nam hendrerit, nunc vitae luctus bibendum, ex elit suscipit
+
        <div class="body-title">Procedure</div>
−
      arcu, eget imperdiet lectus velit in dui. Nunc in ligula a lorem congue volutpat. Etiam at augue sit amet diam
+
            In order to achieve Bronze requirement #4, the Interlab procedure was successfully carried out, following the iGEM 2018 Interlab Study Protocol. To obtain calibration and experimental data, the following materials were used: competent cells from <i>E. coli 's</i> strain DH5α, 96-well plates (black with a clear flat bottom), and the measurement kit, which contains Ludox, silica beads and fluorescein. The first part of the protocol involved generating standard curves for OD measurements, cells and fluorescein. Using a Ludox solution, it was possible to obtain the following reference point for an OD<sub>600</sub>.
−
      pellentesque scelerisque ac a lorem. Pellentesque vel vulputate nulla. Donec lacinia, dolor quis eleifend finibus, orci
+
            The conversion value obtained was then used to convert any measurement of cell density into OD<sub>600</sub> by multiplying by 2.897, using the same instrument under the same conditions.
−
       justo hendrerit mi, et finibus ipsum urna in mi. Sed sollicitudin risus non finibus vehicula. Proin vel rutrum erat.
+
       </div>
−
       Phasellus et lorem mauris. Integer sit amet elit sed risus fermentum mollis eu eu mi. Praesent vestibulum ligula id
+
       <div class="articulo">
−
       purus convallis euismod. Vestibulum dignissim ante id tortor dapibus feugiat non eget ligula. Sed sit amet nisi ac
+
        The particle standard curve was obtained using silica microspheres, which have the same size and optical characteristics as bacteria cells. The standard curve was then used to convert ABS<sub>600</sub> to an estimated number of cells. The following standard curve was obtained:
−
       sapien vehicula lobortis. Suspendisse quis libero at turpis venenatis bibendum sit amet at purus. Aliquam interdum puru
+
       </div>
−
      non mollis molestie. Mauris quis felis vitae neque ornare commodo at id sapien. Vestibulum mattis id libero at suscipit
+
       <div class="imagen-centrada-reducida">
−
    </div>
+
        <img src="https://static.igem.org/mediawiki/2018/8/8b/T--Tec-Monterrey--Interlab_Graph_Abs1.png">
−
    <div class="introduccion">
+
        <div class="leyenda">Figure 1: Calibration curve of particle count from Absorbance at 600 nm.</div>
−
      Lorem ipsum dolor sit amet, consectetur adipiscing elit. Nam hendrerit, nunc vitae luctus bibendum, ex elit suscipit
+
      </div>
−
      arcu, eget imperdiet lectus velit in dui. Nunc in ligula a lorem congue volutpat. Etiam at augue sit amet diam
+
 
−
       pellentesque scelerisque ac a lorem. Pellentesque vel vulputate nulla. Donec lacinia, dolor quis eleifend finibus, orci
+
      <div class="articulo">
−
       justo hendrerit mi, et finibus ipsum urna in mi. Sed sollicitudin risus non finibus vehicula. Proin vel rutrum erat.  
+
        Finally, a fluorescein standard curve was made to calculate the fluorescence values of transformed bacteria with GFP. The filter used for the measurements was used according to the recommended values (Bandpass: 530 nm/ 30 nm, width: 25-30 nm, excitation 485 nm and emission 520-530 nm). The curve displayed in Figure 2 was obtained:
−
       Phasellus et lorem mauris. Integer sit amet elit sed risus fermentum mollis eu eu mi. Praesent vestibulum ligula id
+
       </div>
−
       purus convallis euismod. Vestibulum dignissim ante id tortor dapibus feugiat non eget ligula. Sed sit amet nisi ac
+
       <div class="imagen-centrada-reducida">
−
       sapien vehicula lobortis. Suspendisse quis libero at turpis venenatis bibendum sit amet at purus. Aliquam interdum puru
+
        <img src="https://static.igem.org/mediawiki/2018/3/3a/T--Tec-Monterrey--Interlab_Graph_Fluor1.png">
−
       non mollis molestie. Mauris quis felis vitae neque ornare commodo at id sapien. Vestibulum mattis id libero at suscipit
+
        <div class="leyenda">Figure 2: Calibration curve of Fluorescein</div>
−
    </div>
+
       </div>
−
    <div class="contenido">
+
 
−
    </div>
+
       <div class="contenido">
−
    <div class="contenido_con_imagen_derecha">
+
        <br>
−
    </div>
+
        Using the previous standard curves, it was possible to continue with the cell measurement protocols with transformed <i>E. coli</i> DH5a strains. The following devices from the iGEM kit were transformed:
−
    <div class="contenido_con_imagen_izquierda">
+
        <br>
−
    </div>
+
        <br>
−
    <div class="contenido">
+
       </div>
−
    </div>
+
       <div class="imagen-centrada-reducida">
−
    <div class="imagen_centrada_reducida">
+
        <img src="https://static.igem.org/mediawiki/2018/e/ec/T--Tec-Monterrey--Interlab_Devices.jpeg">
−
    </div>
+
        <div class="leyenda">Figure 3: Devices used to transform DH5α for fluorescence standardising.</div>
−
    <div class="contenido">
+
      </div>
−
    </div>
+
    </section>
−
    <div class="imagen_centrada_completa">
+
 
−
    </div>
+
 
−
    <div class="conclusion">
+
    <section id="results-interlab" class="seccion-responsiva">
−
    </div>
+
      <div class="body-title">Results</div>
−
     <div class="referencias">
+
      <div class="contenido">
−
    </div>
+
        <div class="body-subtitle">Absorbance</div>
−
  </section>
+
        Absorbance of cells at t=0 h and t=6 h are shown in the diagram of Figure 4.
 +
      </div>
 +
      <div class="imagen-centrada-reducida">
 +
        <img src="https://static.igem.org/mediawiki/2018/b/b3/T--Tec-Monterrey--Interlab_Graph_Abs2.png">
 +
        <div class="leyenda">Figure 4: Absorbance at 600 nm of transformed bacteria with devices from iGEM kit.</div>
 +
      </div>
 +
      <div class="contenido">
 +
        <br>
 +
All transformed bacteria show growth after 6 hours. Some of the devices, such as device 1 and 5, did not grow sufficiently as other transformed bacteria did.
 +
      </div>
 +
      <div class="contenido">
 +
        <div class="body-subtitle">Fluorescence</div>
 +
          Fluorescence values at t = 0 h and t = 6 h
 +
      </div>
 +
      <div class="imagen-centrada-reducida">
 +
        <img src="https://static.igem.org/mediawiki/2018/a/a5/T--Tec-Monterrey--Interlab_Graph_Fluor2.png">
 +
        <div class="leyenda">Figure 5: Fluorescence of transformed bacteria with devices from iGEM kit.</div>
 +
      </div>
 +
     </section>
 +
 
 +
    <section id="conclusion-interLab" class="seccion-responsiva">
 +
      <div class="conclusion">
 +
        <div class="body-title">Conclusion</div>
 +
        Fluorescence values obtained by GFP show that some devices emit greater fluorescence values than others. The negative device show no to little fluorescence at both times. Device 3 did not show any fluorescence at all, which affected its measurements. The device that shows the greater fluorescence value was device 4.<br>
 +
        From absorbance and fluorescence values it is evident that fluorescence is not directly related to the culture's optical density, which is to say that absolute GFP expression is not directly related to the amount of bacteria, as it depends on each device. It is then possible to standardise each of these GFP devices, comparing the results from the iGEM Teams that carried out this InterLab protocol, so that amount of bacteria can be related to fluorescence produced by each device.
 +
      </div>
 +
    </section>
 +
 
 +
  </div> <!-- Termina Articulo -->
  
 
</body>
 
</body>
 +
 +
<div class="clear extra_space"></div>
 +
<div class="line_divider"></div>
 +
<div class="clear extra_space"></div>
 +
 
</html>
 
</html>
 
{{:Team:Tec-Monterrey/Templates/Tec-Monterrey_Footer}}
 
{{:Team:Tec-Monterrey/Templates/Tec-Monterrey_Footer}}

Latest revision as of 03:25, 18 October 2018

InterLab
Fifth International InterLab Measurement Study
Procedure
In order to achieve Bronze requirement #4, the Interlab procedure was successfully carried out, following the iGEM 2018 Interlab Study Protocol. To obtain calibration and experimental data, the following materials were used: competent cells from E. coli 's strain DH5α, 96-well plates (black with a clear flat bottom), and the measurement kit, which contains Ludox, silica beads and fluorescein. The first part of the protocol involved generating standard curves for OD measurements, cells and fluorescein. Using a Ludox solution, it was possible to obtain the following reference point for an OD600. The conversion value obtained was then used to convert any measurement of cell density into OD600 by multiplying by 2.897, using the same instrument under the same conditions.
The particle standard curve was obtained using silica microspheres, which have the same size and optical characteristics as bacteria cells. The standard curve was then used to convert ABS600 to an estimated number of cells. The following standard curve was obtained:
Figure 1: Calibration curve of particle count from Absorbance at 600 nm.
Finally, a fluorescein standard curve was made to calculate the fluorescence values of transformed bacteria with GFP. The filter used for the measurements was used according to the recommended values (Bandpass: 530 nm/ 30 nm, width: 25-30 nm, excitation 485 nm and emission 520-530 nm). The curve displayed in Figure 2 was obtained:
Figure 2: Calibration curve of Fluorescein

Using the previous standard curves, it was possible to continue with the cell measurement protocols with transformed E. coli DH5a strains. The following devices from the iGEM kit were transformed:

Figure 3: Devices used to transform DH5α for fluorescence standardising.
Results
Absorbance
Absorbance of cells at t=0 h and t=6 h are shown in the diagram of Figure 4.
Figure 4: Absorbance at 600 nm of transformed bacteria with devices from iGEM kit.

All transformed bacteria show growth after 6 hours. Some of the devices, such as device 1 and 5, did not grow sufficiently as other transformed bacteria did.
Fluorescence
Fluorescence values at t = 0 h and t = 6 h
Figure 5: Fluorescence of transformed bacteria with devices from iGEM kit.
Conclusion
Fluorescence values obtained by GFP show that some devices emit greater fluorescence values than others. The negative device show no to little fluorescence at both times. Device 3 did not show any fluorescence at all, which affected its measurements. The device that shows the greater fluorescence value was device 4.
From absorbance and fluorescence values it is evident that fluorescence is not directly related to the culture's optical density, which is to say that absolute GFP expression is not directly related to the amount of bacteria, as it depends on each device. It is then possible to standardise each of these GFP devices, comparing the results from the iGEM Teams that carried out this InterLab protocol, so that amount of bacteria can be related to fluorescence produced by each device.

Contact Us

Av. Eugenio Garza Sada 2501

64849 Monterrey, Mexico

Sponsors