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{{OUC-China}} | {{OUC-China}} | ||
{{OUC-China/CSS}} | {{OUC-China/CSS}} | ||
+ | <!doctype html> | ||
<html> | <html> | ||
<head> | <head> | ||
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<meta name="viewport" content="width=device-width, initial-scale=1"> | <meta name="viewport" content="width=device-width, initial-scale=1"> | ||
<title>InterLab</title> | <title>InterLab</title> | ||
+ | |||
<style type="text/css"> | <style type="text/css"> | ||
body{padding-top: 50px;} | body{padding-top: 50px;} | ||
− | .ouc-navbar{background-color:#008EA1;top:15px;} | + | .ouc-navbar{background-color:#008EA1; top:15px;} |
.ouc-nav-a{color: white;} | .ouc-nav-a{color: white;} | ||
#ouc-logo{padding-top:5px;} | #ouc-logo{padding-top:5px;} | ||
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.ouc-reserve{background-color: #008EA1;color: white;} | .ouc-reserve{background-color: #008EA1;color: white;} | ||
− | + | .ouc-heading{height:40px;border-left-width:10px;border-left-color:#E3D434; margin-top: 20px;} | |
.ouc-heading h3{margin: 0;color: #008EA1;} | .ouc-heading h3{margin: 0;color: #008EA1;} | ||
.ouc-a{color:#E3D434; } | .ouc-a{color:#E3D434; } | ||
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<body> | <body> | ||
− | + | <!--此处是导航--> | |
− | + | ||
<div class="navbar navbar-fixed-top ouc-navbar" role="navigation"> | <div class="navbar navbar-fixed-top ouc-navbar" role="navigation"> | ||
<ul class="nav navbar-nav"> | <ul class="nav navbar-nav"> | ||
− | <li><a href=" | + | <li><a href="https://2017.igem.org/Team:OUC-China" class="navbar-brand" id="ouc-logo"><img src="https://static.igem.org/mediawiki/2017/8/89/T--OUC-China--logo.jpg" width="66" height="40" /></a></li> |
<li class="dropdown"> | <li class="dropdown"> | ||
− | <a href=" | + | <a href="https://2017.igem.org/Team:OUC-China/Team" data-toggle="dropdown" class="dropdown-toggle ouc-nav-a">Team<span class="caret"></span></a> |
<ul class="dropdown-menu"> | <ul class="dropdown-menu"> | ||
− | <li | + | <li><a href="https://2017.igem.org/Team:OUC-China/Team">Members</a></li> |
<li><a href="#">Collaborations</a></li> | <li><a href="#">Collaborations</a></li> | ||
<li><a href="#">Attributions</a></li> | <li><a href="#">Attributions</a></li> | ||
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</li> | </li> | ||
<li class="dropdown"> | <li class="dropdown"> | ||
− | <a href="#" data-toggle="dropdown" class="dropdown-toggle ouc-nav-a">Project< | + | <a href="#" data-toggle="dropdown" class="dropdown-toggle ouc-nav-a">Project<span class="caret"></span></a> |
<ul class="dropdown-menu"> | <ul class="dropdown-menu"> | ||
<li><a href="#">Description</a></li> | <li><a href="#">Description</a></li> | ||
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<li><a href="#">Experiments</a></li> | <li><a href="#">Experiments</a></li> | ||
<li><a href="#">Results</a></li> | <li><a href="#">Results</a></li> | ||
− | <li><a href=" | + | <li><a href="https://2017.igem.org/Team:OUC-China/InterLab">Interlab</a></li> |
− | <li><a href=" | + | <li><a href="https://2017.igem.org/Team:OUC-China/Notebook" >Notebook</a></li> |
+ | </ul> | ||
+ | </li> | ||
+ | <li class="dropdown"> | ||
+ | <a href="#" data-toggle="dropdown" class="dropdown-toggle ouc-nav-a">Model<span class="caret"></span></a> | ||
+ | <ul class="dropdown-menu"> | ||
+ | <li><a href="#">Overview</a></li> | ||
+ | <li><a href="#">Basic part</a></li> | ||
+ | <li><a href="#">Advanced part</a></li> | ||
</ul> | </ul> | ||
</li> | </li> | ||
− | |||
<li><a href="#" class="ouc-nav-a">Parts</a></li> | <li><a href="#" class="ouc-nav-a">Parts</a></li> | ||
<li><a href="#" class="ouc-nav-a">Safety</a></li> | <li><a href="#" class="ouc-nav-a">Safety</a></li> | ||
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</ul> | </ul> | ||
</div> | </div> | ||
− | + | <!--此处是导航--> | |
<div class="container"> | <div class="container"> | ||
<div class="page-header ouc-page-header"> | <div class="page-header ouc-page-header"> | ||
− | <h1><strong>Interlab</strong></h1> | + | <h1 id="background"><strong>Interlab</strong></h1> |
</div> | </div> | ||
<div class="row"> | <div class="row"> | ||
+ | |||
+ | |||
<div class="col-md-10 "> | <div class="col-md-10 "> | ||
− | <blockquote class="ouc-heading"><h3 | + | |
− | <p> | + | <blockquote class="ouc-heading" ><h3>Background</h3></blockquote> |
+ | <p id="design"> | ||
+ | Fluorescence data can not be directly compared, it is difficult to repeat the measurement. Different instruments, different settings and different measurement environments make this difficult. Through the Interlab study, we expect to have uniform fluorescence measurement steps and standards. These protocols solve this problem by establishing absolute units on the plate reader. | ||
+ | </p> | ||
+ | |||
+ | <blockquote class="ouc-heading"><h3>Design</h3></blockquote> | ||
+ | <p> | ||
+ | <strong>OD 600 Reference point</strong><br/> | ||
+ | Using LUDOX-S40 as a reference, the ratiometric conversion factor of Abs and OD600 can be obtained.<br/> | ||
+ | <strong id="materials">Protocol fluorescein fluorescence standard curve</strong><br/> | ||
+ | By means of the gradient of the fluorescence standard curve, a method of obtaining a fluorescence correction of the cell can be obtained.<br/> | ||
+ | <strong>Cell measurement</strong><br/> | ||
+ | At four time points, the fluorescence intensity of the eight circuits was measured. | ||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
</p> | </p> | ||
− | + | <blockquote class="ouc-heading"><h3>Materials and Method</h3></blockquote> | |
− | <blockquote class="ouc-heading"><h3 | + | |
<p> | <p> | ||
− | <strong> | + | <strong>Plasmids used</strong><br/> |
− | + | Kit Plate 6 InterLab Part Locations<br/> | |
Positive Control (BBa_I20270): well 20B<br/> | Positive Control (BBa_I20270): well 20B<br/> | ||
Negative Control (BBa_R0040): well 20D<br/> | Negative Control (BBa_R0040): well 20D<br/> | ||
− | Test Device 1 (BBa_J364000): well 20F<br/> | + | Test Device 1(BBa_J364000): well 20F<br/> |
− | Test Device 2 (BBa_J364001): well 20H<br/> | + | Test Device 2(BBa_J364001): well 20H<br/> |
− | Test Device 3 (BBa_J364002): well 20J<br/> | + | Test Device 3(BBa_J364002): well 20J<br/> |
− | Test Device 4 (BBa_J364003): well 20L<br/> | + | Test Device 4(BBa_J364003): well 20L<br/> |
− | Test Device 5 (BBa_J364004): well 20N<br/> | + | Test Device 5(BBa_J364004): well 20N<br/> |
− | Test Device 6 (BBa_J364005): well 20P<br/> | + | Test Device 6(BBa_J364005): well 20P<br/> |
− | <strong> | + | <strong>Strain</strong><br/> |
− | E.coli Competent | + | <i>E.coli</i> Competent DH5α ( TaKaRa Code : D9057)<br/> |
− | <strong> | + | <strong>Materials</strong><br/> |
+ | 1ml LUDOX (provided in kit) H20<br/> | ||
+ | Fluorescein (provided in kit) 10ml 1xPBS96 well plate, black with flat bottom<br/> | ||
+ | LB (Luria Bertani) media Chloramphenicol<br/> | ||
+ | 50 ml Falcon tube Incubator at 37°C 1.5 ml eppendorf tubes for sample storage Ice bucket with ice Pipettes<br/> | ||
+ | <strong id="protocol">Machines</strong><br/> | ||
Tecan Spark™ 10M<br/> | Tecan Spark™ 10M<br/> | ||
− | <strong> | + | <strong>Software</strong><br/> |
Microsoft Office EXCEL | Microsoft Office EXCEL | ||
</p> | </p> | ||
− | <blockquote class="ouc-heading"><h3 id=" | + | <blockquote class="ouc-heading"><h3 id="results">Protocol</h3></blockquote> |
− | + | <a href="https://static.igem.org/mediawiki/2017/8/85/InterLab_2017_Plate_Reader_Protocol.pdf">InterLab_2017_Plate_Reader_Protocol</a> | |
− | + | ||
− | + | ||
+ | <blockquote class="ouc-heading"><h3>Results</h3></blockquote> | ||
+ | <strong>OD 600 Reference point</strong><br/> | ||
+ | <table class="table"> | ||
+ | |||
+ | <thead> | ||
+ | |||
+ | <tr> | ||
+ | <th> </th><th>LUDOX-HS40</th><th>H<sub>2</sub>O</th> | ||
+ | </tr> | ||
+ | </thead> | ||
+ | <tbody> | ||
+ | <tr> | ||
+ | <td>Replicate 1</td><td>0.0497</td><td>0.0422</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Replicate 2</td><td>0.0518</td><td>0.0495</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Replicate 3</td><td>0.0546</td><td>0.0428</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Replicate 4</td><td>0.0521</td><td>0.0569</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Arith. Mean</td><td>0.05205</td><td>0.04785</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Corrected Abs600</td><td>0.0042</td><td> </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Reference OD600</td><td>0.0425</td><td> </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>OD600/Abs600</td><td>10.1190476</td><td> </td> | ||
+ | </tr> | ||
+ | </tbody> | ||
+ | |||
+ | </table> | ||
+ | <p style="color: gray;text-align: center;">Table 1. OD600 Reference Point</p> | ||
+ | <br/> | ||
+ | <strong>Protocol fluorescein fluorescence standard curve</strong><br/> | ||
+ | <p style="color: gray;text-align: center;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/8/8e/T--OUC-China--F.S.C_Fluorescein.Standard.Curve.png"><br/> | ||
+ | Figure 1.Fluorescein fluorescence standard curve | ||
+ | </p> | ||
+ | <br/> | ||
+ | <strong>Cell measurement</strong><br/> | ||
+ | <table class="table"> | ||
+ | <thead> | ||
+ | <tr> | ||
+ | <th> </th><th colspan="2" style="text-align: center;">0h</th><th colspan="2" style="text-align: center;">2h</th><th colspan="2" style="text-align: center;">4h</th><th colspan="2" style="text-align: center;">6h</th> | ||
+ | </tr> | ||
+ | </thead> | ||
+ | <tbody> | ||
+ | <tr> | ||
+ | <th> </th><th>Abs600</th><th>FI</th><th>Abs601</th><th>FI</th><th>Abs602</th><th>FI</th><th>Abs603</th><th>FI</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Negative Control (Colony 1)</td><td>0.0301</td><td>7</td><td>0.1156</td><td>8</td><td>0.5106</td><td>14</td><td>0.7453</td><td>17</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Negative Control (Colony 2)</td><td>0.0408</td><td>5</td><td>0.1352</td><td>8</td><td>0.5184</td><td>17</td><td>0.7326</td><td>16</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Positive Control (Colony 1)</td><td>0.0308</td><td>28</td><td>0.0805</td><td>65</td><td>0.4715</td><td>476</td><td>0.7293</td><td>722</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Positive Control (Colony 2)</td><td>0.0437</td><td>31</td><td>0.1498</td><td>136</td><td>0.4956</td><td>428</td><td>0.7049</td><td>677</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 1: J23101.BCD2.E0040.B0015 (Colony 1)</td><td>0.0283</td><td>46</td><td>0.0883</td><td>152</td><td>0.4358</td><td>606</td><td>0.6846</td><td>948</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 1: J23101.BCD2.E0040.B0015 (Colony 2)</td><td>0.0261</td><td>38</td><td>0.0969</td><td>119</td><td>0.3719</td><td>464</td><td>0.5960</td><td>738</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 2: J23106.BCD2.E0040.B0015 (Colony 1)</td><td>0.0378</td><td>44</td><td>0.1120</td><td>164</td><td>0.5090</td><td>718</td><td>0.7145</td><td>1258</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 2: J23106.BCD2.E0040.B0015 (Colony 2)</td><td>0.0223</td><td>16</td><td>0.0788</td><td>115</td><td>0.4045</td><td>506</td><td>0.5945</td><td>963</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 3: J23117.BCD2.E0040.B0015 (Colony 1)</td><td>0.0323</td><td>1</td><td>0.0945</td><td>10</td><td>0.4482</td><td>22</td><td>0.5704</td><td>28</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 3: J23117.BCD2.E0040.B0015 (Colony 2)</td><td>0.0156</td><td>4</td><td>0.0537</td><td>8</td><td>0.3399</td><td>18</td><td>0.5254</td><td>24</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 4: J23101+I13504 (Colony 1)</td><td>0.0201</td><td>9</td><td>0.0585</td><td>76</td><td>0.3224</td><td>339</td><td>0.4975</td><td>472</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 4: J23101+I13504 (Colony 2)</td><td>0.0257</td><td>16</td><td>0.0628</td><td>84</td><td>0.3267</td><td>340</td><td>0.4900</td><td>471</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 5: J23106+I13504 (Colony 1)</td><td>0.0263</td><td>7</td><td>0.0804</td><td>46</td><td>0.4000</td><td>106</td><td>0.5413</td><td>120</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 5: J23106+I13504 (Colony 2)</td><td>0.0269</td><td>6</td><td>0.0850</td><td>45</td><td>0.3789</td><td>94</td><td>0.5285</td><td>111</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 6: J23117+I13504 (Colony 1)</td><td>0.0265</td><td>1</td><td>0.0959</td><td>8</td><td>0.3967</td><td>15</td><td>0.4837</td><td>2</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 6: J23117+I13504 (Colony 2)</td><td>0.0243</td><td>-1</td><td>0.0883</td><td>5</td><td>0.4016</td><td>13</td><td>0.5212</td><td>10</td> | ||
+ | </tr> | ||
+ | </tbody> | ||
+ | </table> | ||
+ | <p style="color: gray;text-align: center;">Table 2. Raw data of Abs600 and Fluorescence measurement</p> | ||
+ | <br/> | ||
+ | <table class="table"> | ||
+ | <thead> | ||
+ | <tr> | ||
+ | <th> </th><th colspan="2" style="text-align: center;">0h</th><th colspan="2" style="text-align: center;">2h</th><th colspan="2" style="text-align: center;">4h</th><th colspan="2" style="text-align: center;">6h</th> | ||
+ | </tr> | ||
+ | </thead> | ||
+ | <tbody> | ||
+ | <tr> | ||
+ | <th> </th><th>average</th><th>SD</th><th>average</th><th>SD</th><th>average</th><th>SD</th><th>average</th><th>SD</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Negative Control (Colony 1)</td><td>232</td><td>60</td><td>75</td><td>28</td><td>27</td><td>6</td><td>23</td><td>6</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Negative Control (Colony 2)</td><td>114</td><td>11</td><td>59</td><td>25</td><td>34</td><td>4</td><td>22</td><td>3</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Positive Control (Colony 1)</td><td>920</td><td>180</td><td>827</td><td>222</td><td>1013</td><td>75</td><td>994</td><td>87</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Positive Control (Colony 2)</td><td>720</td><td>40</td><td>911</td><td>86</td><td>867</td><td>81</td><td>960</td><td>15</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 1: J23101.BCD2.E0040.B0015 (Colony 1)</td><td>1636</td><td>324</td><td>1748</td><td>295</td><td>1394</td><td>80</td><td>1388</td><td>108</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 1: J23101.BCD2.E0040.B0015 (Colony 2)</td><td>1462</td><td>64</td><td>1227</td><td>80</td><td>1248</td><td>64</td><td>1239</td><td>28</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 2: J23106.BCD2.E0040.B0015 (Colony 1)</td><td>1190</td><td>212</td><td>1489</td><td>231</td><td>1413</td><td>69</td><td>1763</td><td>92</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 2: J23106.BCD2.E0040.B0015 (Colony 2)</td><td>716</td><td>103</td><td>1469</td><td>163</td><td>1250</td><td>54</td><td>1621</td><td>83</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 3: J23117.BCD2.E0040.B0015 (Colony 1)</td><td>27</td><td>61</td><td>112</td><td>50</td><td>49</td><td>7</td><td>49</td><td>11</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 3: J23117.BCD2.E0040.B0015 (Colony 2)</td><td>282</td><td>108</td><td>148</td><td>45</td><td>52</td><td>3</td><td>45</td><td>11</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 4: J23101+I13504 (Colony 1)</td><td>461</td><td>76</td><td>1319</td><td>170</td><td>1052</td><td>27</td><td>947</td><td>86</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 4: J23101+I13504 (Colony 2)</td><td>617</td><td>136</td><td>1350</td><td>189</td><td>1042</td><td>73</td><td>962</td><td>51</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 5: J23106+I13504 (Colony 1)</td><td>280</td><td>175</td><td>576</td><td>77</td><td>266</td><td>16</td><td>221</td><td>14</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 5: J23106+I13504 (Colony 2)</td><td>215</td><td>93</td><td>530</td><td>41</td><td>249</td><td>20</td><td>211</td><td>12</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 6: J23117+I13504 (Colony 1)</td><td>34</td><td>73</td><td>78</td><td>41</td><td>38</td><td>8</td><td>4</td><td>8</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Test Device 6: J23117+I13504 (Colony 2)</td><td>-55</td><td>102</td><td>60</td><td>23</td><td>33</td><td>3</td><td>20</td><td>4</td> | ||
+ | </tr> | ||
+ | </tbody> | ||
+ | </table> | ||
+ | <p style="color: gray;text-align: center;">Table 3. Data of Fluorescence/Abs600</p> | ||
+ | <p style="color: gray;text-align: center;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/4/4d/T--OUC-China--FI_Abs600_for_8_devices.FI_Abs%EF%BC%88AU%EF%BC%89.png" class="img-thumbnail"><br/> | ||
+ | Figure 2.Average level of devices | ||
+ | </p> | ||
<blockquote class="ouc-heading"><h3 id="discussion">Discussion</h3></blockquote> | <blockquote class="ouc-heading"><h3 id="discussion">Discussion</h3></blockquote> | ||
+ | <p>Device 2 and device 3 have strong fluorescence. Device 4 and device 5 have moderate fluorescence intensity. The device 3 and the device 6 are weakly fluorescent.</p> | ||
</div> | </div> | ||
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<div class="col-md-2 "> | <div class="col-md-2 "> | ||
<ul class="nav nav-stacked affix ouc-ul"> | <ul class="nav nav-stacked affix ouc-ul"> | ||
− | |||
<li><a href="#background" class="ouc-a">Background</a></li> | <li><a href="#background" class="ouc-a">Background</a></li> | ||
− | <li><a href="#materials" class="ouc-a">Materials</a></li> | + | <li><a href="#design" class="ouc-a">Design</a></li> |
+ | <li><a href="#materials" class="ouc-a">Materials and Method</a></li> | ||
<li><a href="#protocol" class="ouc-a">Protocol</a></li> | <li><a href="#protocol" class="ouc-a">Protocol</a></li> | ||
<li><a href="#results" class="ouc-a">Results</a></li> | <li><a href="#results" class="ouc-a">Results</a></li> | ||
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</div> | </div> | ||
</div> | </div> | ||
− | + | ||
− | + | ||
+ | <!--此处是置顶及赞助页--> | ||
<a href="#top"><img class="top" src="https://static.igem.org/mediawiki/2017/b/b7/T--OUC-China--top.jpeg" width="101" height="101" /></a> | <a href="#top"><img class="top" src="https://static.igem.org/mediawiki/2017/b/b7/T--OUC-China--top.jpeg" width="101" height="101" /></a> | ||
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Revision as of 10:48, 23 September 2017
Interlab
Background
Fluorescence data can not be directly compared, it is difficult to repeat the measurement. Different instruments, different settings and different measurement environments make this difficult. Through the Interlab study, we expect to have uniform fluorescence measurement steps and standards. These protocols solve this problem by establishing absolute units on the plate reader.
Design
OD 600 Reference point
Using LUDOX-S40 as a reference, the ratiometric conversion factor of Abs and OD600 can be obtained.
Protocol fluorescein fluorescence standard curve
By means of the gradient of the fluorescence standard curve, a method of obtaining a fluorescence correction of the cell can be obtained.
Cell measurement
At four time points, the fluorescence intensity of the eight circuits was measured.
Materials and Method
Plasmids used
Kit Plate 6 InterLab Part Locations
Positive Control (BBa_I20270): well 20B
Negative Control (BBa_R0040): well 20D
Test Device 1(BBa_J364000): well 20F
Test Device 2(BBa_J364001): well 20H
Test Device 3(BBa_J364002): well 20J
Test Device 4(BBa_J364003): well 20L
Test Device 5(BBa_J364004): well 20N
Test Device 6(BBa_J364005): well 20P
Strain
E.coli Competent DH5α ( TaKaRa Code : D9057)
Materials
1ml LUDOX (provided in kit) H20
Fluorescein (provided in kit) 10ml 1xPBS96 well plate, black with flat bottom
LB (Luria Bertani) media Chloramphenicol
50 ml Falcon tube Incubator at 37°C 1.5 ml eppendorf tubes for sample storage Ice bucket with ice Pipettes
Machines
Tecan Spark™ 10M
Software
Microsoft Office EXCEL
InterLab_2017_Plate_Reader_ProtocolProtocol
OD 600 Reference pointResults
LUDOX-HS40 | H2O | |
---|---|---|
Replicate 1 | 0.0497 | 0.0422 |
Replicate 2 | 0.0518 | 0.0495 |
Replicate 3 | 0.0546 | 0.0428 |
Replicate 4 | 0.0521 | 0.0569 |
Arith. Mean | 0.05205 | 0.04785 |
Corrected Abs600 | 0.0042 | |
Reference OD600 | 0.0425 | |
OD600/Abs600 | 10.1190476 |
Table 1. OD600 Reference Point
Protocol fluorescein fluorescence standard curve
Figure 1.Fluorescein fluorescence standard curve
Cell measurement
0h | 2h | 4h | 6h | |||||
---|---|---|---|---|---|---|---|---|
Abs600 | FI | Abs601 | FI | Abs602 | FI | Abs603 | FI | |
Negative Control (Colony 1) | 0.0301 | 7 | 0.1156 | 8 | 0.5106 | 14 | 0.7453 | 17 |
Negative Control (Colony 2) | 0.0408 | 5 | 0.1352 | 8 | 0.5184 | 17 | 0.7326 | 16 |
Positive Control (Colony 1) | 0.0308 | 28 | 0.0805 | 65 | 0.4715 | 476 | 0.7293 | 722 |
Positive Control (Colony 2) | 0.0437 | 31 | 0.1498 | 136 | 0.4956 | 428 | 0.7049 | 677 |
Test Device 1: J23101.BCD2.E0040.B0015 (Colony 1) | 0.0283 | 46 | 0.0883 | 152 | 0.4358 | 606 | 0.6846 | 948 |
Test Device 1: J23101.BCD2.E0040.B0015 (Colony 2) | 0.0261 | 38 | 0.0969 | 119 | 0.3719 | 464 | 0.5960 | 738 |
Test Device 2: J23106.BCD2.E0040.B0015 (Colony 1) | 0.0378 | 44 | 0.1120 | 164 | 0.5090 | 718 | 0.7145 | 1258 |
Test Device 2: J23106.BCD2.E0040.B0015 (Colony 2) | 0.0223 | 16 | 0.0788 | 115 | 0.4045 | 506 | 0.5945 | 963 |
Test Device 3: J23117.BCD2.E0040.B0015 (Colony 1) | 0.0323 | 1 | 0.0945 | 10 | 0.4482 | 22 | 0.5704 | 28 |
Test Device 3: J23117.BCD2.E0040.B0015 (Colony 2) | 0.0156 | 4 | 0.0537 | 8 | 0.3399 | 18 | 0.5254 | 24 |
Test Device 4: J23101+I13504 (Colony 1) | 0.0201 | 9 | 0.0585 | 76 | 0.3224 | 339 | 0.4975 | 472 |
Test Device 4: J23101+I13504 (Colony 2) | 0.0257 | 16 | 0.0628 | 84 | 0.3267 | 340 | 0.4900 | 471 |
Test Device 5: J23106+I13504 (Colony 1) | 0.0263 | 7 | 0.0804 | 46 | 0.4000 | 106 | 0.5413 | 120 |
Test Device 5: J23106+I13504 (Colony 2) | 0.0269 | 6 | 0.0850 | 45 | 0.3789 | 94 | 0.5285 | 111 |
Test Device 6: J23117+I13504 (Colony 1) | 0.0265 | 1 | 0.0959 | 8 | 0.3967 | 15 | 0.4837 | 2 |
Test Device 6: J23117+I13504 (Colony 2) | 0.0243 | -1 | 0.0883 | 5 | 0.4016 | 13 | 0.5212 | 10 |
Table 2. Raw data of Abs600 and Fluorescence measurement
0h | 2h | 4h | 6h | |||||
---|---|---|---|---|---|---|---|---|
average | SD | average | SD | average | SD | average | SD | |
Negative Control (Colony 1) | 232 | 60 | 75 | 28 | 27 | 6 | 23 | 6 |
Negative Control (Colony 2) | 114 | 11 | 59 | 25 | 34 | 4 | 22 | 3 |
Positive Control (Colony 1) | 920 | 180 | 827 | 222 | 1013 | 75 | 994 | 87 |
Positive Control (Colony 2) | 720 | 40 | 911 | 86 | 867 | 81 | 960 | 15 |
Test Device 1: J23101.BCD2.E0040.B0015 (Colony 1) | 1636 | 324 | 1748 | 295 | 1394 | 80 | 1388 | 108 |
Test Device 1: J23101.BCD2.E0040.B0015 (Colony 2) | 1462 | 64 | 1227 | 80 | 1248 | 64 | 1239 | 28 |
Test Device 2: J23106.BCD2.E0040.B0015 (Colony 1) | 1190 | 212 | 1489 | 231 | 1413 | 69 | 1763 | 92 |
Test Device 2: J23106.BCD2.E0040.B0015 (Colony 2) | 716 | 103 | 1469 | 163 | 1250 | 54 | 1621 | 83 |
Test Device 3: J23117.BCD2.E0040.B0015 (Colony 1) | 27 | 61 | 112 | 50 | 49 | 7 | 49 | 11 |
Test Device 3: J23117.BCD2.E0040.B0015 (Colony 2) | 282 | 108 | 148 | 45 | 52 | 3 | 45 | 11 |
Test Device 4: J23101+I13504 (Colony 1) | 461 | 76 | 1319 | 170 | 1052 | 27 | 947 | 86 |
Test Device 4: J23101+I13504 (Colony 2) | 617 | 136 | 1350 | 189 | 1042 | 73 | 962 | 51 |
Test Device 5: J23106+I13504 (Colony 1) | 280 | 175 | 576 | 77 | 266 | 16 | 221 | 14 |
Test Device 5: J23106+I13504 (Colony 2) | 215 | 93 | 530 | 41 | 249 | 20 | 211 | 12 |
Test Device 6: J23117+I13504 (Colony 1) | 34 | 73 | 78 | 41 | 38 | 8 | 4 | 8 |
Test Device 6: J23117+I13504 (Colony 2) | -55 | 102 | 60 | 23 | 33 | 3 | 20 | 4 |
Table 3. Data of Fluorescence/Abs600
Figure 2.Average level of devices
Discussion
Device 2 and device 3 have strong fluorescence. Device 4 and device 5 have moderate fluorescence intensity. The device 3 and the device 6 are weakly fluorescent.
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