Difference between revisions of "Team:Aix-Marseille/Notebook"

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__NOTOC__
 
__NOTOC__
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{{:Team:Aix-Marseille/Template-Style-and-Menu}}<html>
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{{:Team:Aix-Marseille/Template-Style-and-Menu}}{{:Team:Aix-Marseille/Template-Notebook}}<div class="events">
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<script> // Unhides and colours the calendar :)
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$(function() {
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$(".day time").each(function(index) {
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adate = "#" + $(this).attr("datetime");
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−
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if ($(adate).length) {
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n = $(adate + " > ul > li").length;
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−
 
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if      (n >= 8) c = "5";
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else if (n >= 6) c = "4";
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else if (n >= 4) c = "3";
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else if (n >= 3) c = "2";
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else if (n >= 2) c = "1";
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else            c = "0";
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$(this).addClass("day-"+c);
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$(this).children("a").attr("href", adate);
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}
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});
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$(".calendar").removeClass("hide");
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        $(".evday").hide();
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});
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</script>
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<style>
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.calendar { display: flex;
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flex-wrap: wrap;
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font-family: "Raleway", sans-serif;
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}
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.calendar.hide { display: none; }
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.evday:target { display: block !important; }
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.evday { margin: 3em; }
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.events { flex: 1 }
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.week {
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  display: flex;
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  list-style-type: none;
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  margin: 0;
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  padding: 0;
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}
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.day {
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  width: calc(100% / 7 - 1em);
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  min-width: 2em;
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  min-height: 2em;
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  margin: .5em;
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  display: block;
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}
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−
 
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.day a[href]:hover {
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  transition: all 100ms ease-in;
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−
  transform: scale(1.2);
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  font-weight: bold;
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}
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.week:first-of-type {
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  justify-content: flex-end;
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}
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.week:last-of-type {
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  justify-content: flex-start;
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}
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.month {
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  max-width: 460px;
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  margin: 20px auto;
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}
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.day a {
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  display: block;
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  text-shadow: 2px 2px rgba(210,83,65,0.4);
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  box-shadow: 2px 2px rgba(210,83,65,0.4);
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  transition: all 50ms ease-out;
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−
  background-color: #fff;
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  padding: 5px;
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  text-decoration: none;
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  color: #fff;
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}
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time.day-0 a { background-color: #ffeeeb; }
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time.day-1 a { background-color: #ffccc4; }
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time.day-2 a { background-color: #ffa99d; }
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time.day-3 a { background-color: #ff8776; }
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time.day-4 a { background-color: #ff654e; }
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time.day-5 a { background-color: #ff3114; }
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</style>
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<div class="content_wrapper" style="display: flex; flex-direction: column;">
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<div style="background-color: #fff; flex: 1;">
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<div class="calendar hide">
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−
  <div class="month">
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−
  <h2>June</h2>
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−
    <ul class="week">
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−
      <li class="day"><time datetime="2016-06-01"><a>1</a></time></li>
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−
      <li class="day"><time datetime="2016-06-02"><a>2</a></time></li>
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−
      <li class="day"><time datetime="2016-06-03"><a>3</a></time></li>
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−
      <li class="day"><time datetime="2016-06-04"><a>4</a></time></li>
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−
      <li class="day"><time datetime="2016-06-05"><a>5</a></time></li>
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    </ul>
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−
    <ul class="week">
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−
      <li class="day"><time datetime="2016-06-06"><a>6</a></time></li>
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−
      <li class="day"><time datetime="2016-06-07"><a>7</a></time></li>
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−
      <li class="day"><time datetime="2016-06-08"><a>8</a></time></li>
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−
      <li class="day"><time datetime="2016-06-09"><a>9</a></time></li>
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−
      <li class="day"><time datetime="2016-06-10"><a>10</a></time></li>
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−
      <li class="day"><time datetime="2016-06-11"><a>11</a></time></li>
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      <li class="day"><time datetime="2016-06-12"><a>12</a></time></li>
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−
    </ul>
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−
    <ul class="week">
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−
      <li class="day"><time datetime="2016-06-13"><a>13</a></time></li>
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      <li class="day"><time datetime="2016-06-14"><a>14</a></time></li>
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−
      <li class="day"><time datetime="2016-06-15"><a>15</a></time></li>
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−
      <li class="day"><time datetime="2016-06-16"><a>16</a></time></li>
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−
      <li class="day"><time datetime="2016-06-17"><a>17</a></time></li>
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−
      <li class="day"><time datetime="2016-06-18"><a>18</a></time></li>
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−
      <li class="day"><time datetime="2016-06-19"><a>19</a></time></li>
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−
    </ul>
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−
    <ul class="week">
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−
      <li class="day"><time datetime="2016-06-20"><a>20</a></time></li>
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−
      <li class="day"><time datetime="2016-06-21"><a>21</a></time></li>
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−
      <li class="day"><time datetime="2016-06-22"><a>22</a></time></li>
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−
      <li class="day"><time datetime="2016-06-23"><a>23</a></time></li>
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−
      <li class="day"><time datetime="2016-06-24"><a>24</a></time></li>
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−
      <li class="day"><time datetime="2016-06-25"><a>25</a></time></li>
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−
      <li class="day"><time datetime="2016-06-26"><a>26</a></time></li>
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−
    </ul>
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−
    <ul class="week">
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−
      <li class="day"><time datetime="2016-06-27"><a>27</a></time></li>
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−
      <li class="day"><time datetime="2016-06-28"><a>28</a></time></li>
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−
      <li class="day"><time datetime="2016-06-29"><a>29</a></time></li>
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−
      <li class="day"><time datetime="2016-06-30"><a>30</a></time></li>
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−
    </ul>
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−
  </div>
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−
 
+
−
 
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−
  <div class="month">
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−
  <h2>July</h2>
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−
    <ul class="week">
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−
      <li class="day"><time datetime="2016-07-01"><a>1</a></time></li>
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−
      <li class="day"><time datetime="2016-07-02"><a>2</a></time></li>
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−
      <li class="day"><time datetime="2016-07-03"><a>3</a></time></li>
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−
    </ul>
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−
    <ul class="week">
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−
      <li class="day"><time datetime="2016-07-04"><a>4</a></time></li>
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−
      <li class="day"><time datetime="2016-07-05"><a>5</a></time></li>
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−
      <li class="day"><time datetime="2016-07-06"><a>6</a></time></li>
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−
      <li class="day"><time datetime="2016-07-07"><a>7</a></time></li>
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−
      <li class="day"><time datetime="2016-07-08"><a>8</a></time></li>
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−
      <li class="day"><time datetime="2016-07-09"><a>9</a></time></li>
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−
      <li class="day"><time datetime="2016-07-10"><a>10</a></time></li>
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−
    </ul>
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−
    <ul class="week">
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−
      <li class="day"><time datetime="2016-07-11"><a>11</a></time></li>
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−
      <li class="day"><time datetime="2016-07-12"><a>12</a></time></li>
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−
      <li class="day"><time datetime="2016-07-13"><a>13</a></time></li>
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−
      <li class="day"><time datetime="2016-07-14"><a>14</a></time></li>
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−
      <li class="day"><time datetime="2016-07-15"><a>15</a></time></li>
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−
      <li class="day"><time datetime="2016-07-16"><a>16</a></time></li>
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−
      <li class="day"><time datetime="2016-07-17"><a>17</a></time></li>
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−
 
+
−
    </ul>
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−
    <ul class="week">
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−
      <li class="day"><time datetime="2016-07-18"><a>18</a></time></li>
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−
      <li class="day"><time datetime="2016-07-19"><a>19</a></time></li>
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−
      <li class="day"><time datetime="2016-07-20"><a>20</a></time></li>
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−
      <li class="day"><time datetime="2016-07-21"><a>21</a></time></li>
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−
      <li class="day"><time datetime="2016-07-22"><a>22</a></time></li>
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−
      <li class="day"><time datetime="2016-07-23"><a>23</a></time></li>
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−
      <li class="day"><time datetime="2016-07-24"><a>24</a></time></li>
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−
    </ul>
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−
    <ul class="week">
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−
      <li class="day"><time datetime="2016-07-25"><a>25</a></time></li>
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−
      <li class="day"><time datetime="2016-07-26"><a>26</a></time></li>
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−
      <li class="day"><time datetime="2016-07-27"><a>27</a></time></li>
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−
      <li class="day"><time datetime="2016-07-28"><a>28</a></time></li>
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−
      <li class="day"><time datetime="2016-07-29"><a>29</a></time></li>
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−
      <li class="day"><time datetime="2016-07-30"><a>30</a></time></li>
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−
      <li class="day"><time datetime="2016-07-31"><a>31</a></time></li>
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−
    </ul>
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−
  </div>
+
−
 
+
−
  <div class="month">
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−
  <h2>August</h2>
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−
    <ul class="week">
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−
      <li class="day"><time datetime="2016-08-01"><a>1</a></time></li>
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−
      <li class="day"><time datetime="2016-08-02"><a>2</a></time></li>
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−
      <li class="day"><time datetime="2016-08-03"><a>3</a></time></li>
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−
      <li class="day"><time datetime="2016-08-04"><a>4</a></time></li>
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−
      <li class="day"><time datetime="2016-08-05"><a>5</a></time></li>
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−
      <li class="day"><time datetime="2016-08-06"><a>6</a></time></li>
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−
      <li class="day"><time datetime="2016-08-07"><a>7</a></time></li>
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−
    </ul>
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−
    <ul class="week">
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−
      <li class="day"><time datetime="2016-08-08"><a>8</a></time></li>
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      <li class="day"><time datetime="2016-08-09"><a>9</a></time></li>
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−
      <li class="day"><time datetime="2016-08-10"><a>10</a></time></li>
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−
      <li class="day"><time datetime="2016-08-11"><a>11</a></time></li>
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−
      <li class="day"><time datetime="2016-08-12"><a>12</a></time></li>
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−
      <li class="day"><time datetime="2016-08-13"><a>13</a></time></li>
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−
      <li class="day"><time datetime="2016-08-14"><a>14</a></time></li>
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−
    </ul>
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−
    <ul class="week">
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−
      <li class="day"><time datetime="2016-08-15"><a>15</a></time></li>
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−
      <li class="day"><time datetime="2016-08-16"><a>16</a></time></li>
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−
      <li class="day"><time datetime="2016-08-17"><a>17</a></time></li>
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−
      <li class="day"><time datetime="2016-08-18"><a>18</a></time></li>
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−
      <li class="day"><time datetime="2016-08-19"><a>19</a></time></li>
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−
      <li class="day"><time datetime="2016-08-20"><a>20</a></time></li>
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−
      <li class="day"><time datetime="2016-08-21"><a>21</a></time></li>
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−
    </ul>
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−
    <ul class="week">
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−
      <li class="day"><time datetime="2016-08-22"><a>22</a></time></li>
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−
      <li class="day"><time datetime="2016-08-23"><a>23</a></time></li>
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−
      <li class="day"><time datetime="2016-08-24"><a>24</a></time></li>
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−
      <li class="day"><time datetime="2016-08-25"><a>25</a></time></li>
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−
      <li class="day"><time datetime="2016-08-26"><a>26</a></time></li>
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−
      <li class="day"><time datetime="2016-08-27"><a>27</a></time></li>
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−
      <li class="day"><time datetime="2016-08-28"><a>28</a></time></li>
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−
    </ul>
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−
    <ul class="week">
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−
      <li class="day"><time datetime="2016-08-29"><a>29</a></time></li>
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−
      <li class="day"><time datetime="2016-08-30"><a>30</a></time></li>
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−
      <li class="day"><time datetime="2016-08-31"><a>31</a></time></li>
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−
    </ul>
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−
  </div>
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−
  <div class="month">
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−
  <h2>September</h2>
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−
    <ul class="week">
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−
      <li class="day"><time datetime="2016-09-01"><a>1</a></time></li>
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−
      <li class="day"><time datetime="2016-09-02"><a>2</a></time></li>
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−
      <li class="day"><time datetime="2016-09-03"><a>3</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-04"><a>4</a></time></li>
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−
    </ul>
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−
    <ul class="week">
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−
      <li class="day"><time datetime="2016-09-05"><a>5</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-06"><a>6</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-07"><a>7</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-08"><a>8</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-09"><a>9</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-10"><a>10</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-11"><a>11</a></time></li>
+
−
    </ul>
+
−
    <ul class="week">
+
−
      <li class="day"><time datetime="2016-09-12"><a>12</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-13"><a>13</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-14"><a>14</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-15"><a>15</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-16"><a>16</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-17"><a>17</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-18"><a>18</a></time></li>
+
−
    </ul>
+
−
    <ul class="week">
+
−
      <li class="day"><time datetime="2016-09-19"><a>19</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-20"><a>20</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-21"><a>21</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-22"><a>22</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-23"><a>23</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-24"><a>24</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-25"><a>25</a></time></li>
+
−
    </ul>
+
−
    <ul class="week">
+
−
      <li class="day"><time datetime="2016-09-26"><a>26</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-27"><a>27</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-28"><a>28</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-29"><a>29</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-30"><a>30</a></time></li>
+
−
      <li class="day"><time datetime="2016-09-31"><a>31</a></time></li>
+
−
    </ul>
+
−
  </div>
+
−
</div></html><div class="events">
+
 
<div id="2016-06-13" class="evday">
 
<div id="2016-06-13" class="evday">
  
Line 552: Line 282:
 
* Colony PCR of CsgA from the 01/08 using EconoTac ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).
 
* Colony PCR of CsgA from the 01/08 using EconoTac ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).
 
* PCR clean up of DesC(200ng/µL) and gel extraction of DesA (83ng/µL), DesB(53ng/µL), FliC <i>E. coli</i> ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.237_:_PCR_clean-up_and_gel_extraction Protocol#7]).
 
* PCR clean up of DesC(200ng/µL) and gel extraction of DesA (83ng/µL), DesB(53ng/µL), FliC <i>E. coli</i> ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.237_:_PCR_clean-up_and_gel_extraction Protocol#7]).
−
* SLIC of DesA, DesB, DesC, DesD and transformation of 10µL in 100µL competant Tg1 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#.23Protocol_7_:_SLIC_.28sequence-_and_ligation-independent_cloning.29 Protocol#10]).
+
* SLIC of DesA, DesB, DesC, DesD and transformation of 10µL in 100µL competent Tg1 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#.23Protocol_7_:_SLIC_.28sequence-_and_ligation-independent_cloning.29 Protocol#10]).
 
* Sequencage of Desulfovribrio clone 3 validated.
 
* Sequencage of Desulfovribrio clone 3 validated.
 
* E/P digestion on pSB1C3, pSB1A3 and pSB1K3 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.235_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
 
* E/P digestion on pSB1C3, pSB1A3 and pSB1K3 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.235_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
Line 586: Line 316:
 
[[File:T--Aix-Marseille--PCR-08-08.jpg|left|thumb|Colony PCR verification. The clones picked up on the plates are wroten on the right of the gel. the awaited size is 506bp for all clones.]]
 
[[File:T--Aix-Marseille--PCR-08-08.jpg|left|thumb|Colony PCR verification. The clones picked up on the plates are wroten on the right of the gel. the awaited size is 506bp for all clones.]]
 
* Ligation of BBa_B0034+BBa_I0500 and Bba_K1951000 into pSB1K3 to create desA producer ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
 
* Ligation of BBa_B0034+BBa_I0500 and Bba_K1951000 into pSB1K3 to create desA producer ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
−
* Transformation of the previous ligation in competant Tg1  ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations protocol#2]).
+
* Transformation of the previous ligation in competent Tg1  ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations protocol#2]).
 
* Starters from DesC and Bba_K1951008 of the 08.04.2016.
 
* Starters from DesC and Bba_K1951008 of the 08.04.2016.
 
* Resuspension of oligo using the IDT protocole.
 
* Resuspension of oligo using the IDT protocole.
Line 671: Line 401:
 
* PCR using Q5 master with new FliC coli sequence (substitute Bbs1 site) Bba_K1951005 and Bba_K1951001, every SLIC have been validated by electrophoresis ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).
 
* PCR using Q5 master with new FliC coli sequence (substitute Bbs1 site) Bba_K1951005 and Bba_K1951001, every SLIC have been validated by electrophoresis ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).
 
* PCR clean up on the 2 previous SLIC  ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.237_:_PCR_clean-up_and_gel_extraction Protocol#7]).
 
* PCR clean up on the 2 previous SLIC  ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.237_:_PCR_clean-up_and_gel_extraction Protocol#7]).
−
* SLIC and Transformation of Bba_K1951005 and Bba_K1951001 in competant Tg1 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#.23Protocol_7_:_SLIC_.28sequence-_and_ligation-independent_cloning.29 Protocol#10]).
+
* SLIC and Transformation of Bba_K1951005 and Bba_K1951001 in competent Tg1 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#.23Protocol_7_:_SLIC_.28sequence-_and_ligation-independent_cloning.29 Protocol#10]).
 
* Starter of Bba_K1951004, Bba_K1951009 and Bba_K1951010 to make a protein production test.
 
* Starter of Bba_K1951004, Bba_K1951009 and Bba_K1951010 to make a protein production test.
 
</div>
 
</div>
Line 722: Line 452:
 
* Pcr clean up on the digestion from the 08.25.2016 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.237_:_PCR_clean-up_and_gel_extraction Protocol#7]).
 
* Pcr clean up on the digestion from the 08.25.2016 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.237_:_PCR_clean-up_and_gel_extraction Protocol#7]).
 
* Ligation using 2A method of Bba_K1951002 or Bba_K1951003 and Bba_B0034 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
 
* Ligation using 2A method of Bba_K1951002 or Bba_K1951003 and Bba_B0034 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
−
* Transformation of the previous ligation in competant Tg1 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
+
* Transformation of the previous ligation in competent Tg1 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
 
* Digestion of 500ng in 50µL total mix of Bba_K1951002 and Bba_K1951005 by XbaI and PstI ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
 
* Digestion of 500ng in 50µL total mix of Bba_K1951002 and Bba_K1951005 by XbaI and PstI ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
 
* Ligation of Bba_K1951001(DesB), Bba_K1951002(DesC), Bba_K1951003(DesD) and Bba_B0034 (rbs) overnight at 16°C ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
 
* Ligation of Bba_K1951001(DesB), Bba_K1951002(DesC), Bba_K1951003(DesD) and Bba_B0034 (rbs) overnight at 16°C ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
Line 742: Line 472:
 
* Colony PCR of Bba_K1951001(DesB), Bba_K1951002(DesC), Bba_K1951003(DesD) and Bba_B0034 (rbs) ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).
 
* Colony PCR of Bba_K1951001(DesB), Bba_K1951002(DesC), Bba_K1951003(DesD) and Bba_B0034 (rbs) ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).
 
* Colony PCR of Bba_K1951001(DesB), Bba_K1951002(DesC), Bba_K1951003(DesD) and Bba_B0034 (rbs) + BBa_I0500 (Prom) ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).
 
* Colony PCR of Bba_K1951001(DesB), Bba_K1951002(DesC), Bba_K1951003(DesD) and Bba_B0034 (rbs) + BBa_I0500 (Prom) ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).
−
* Transformation of Bba_K1951004 (DesA with pSB1C3) in competant Tg1 cells ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
+
* Transformation of Bba_K1951004 (DesA with pSB1C3) in competent Tg1 cells ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
 
* Ligation of Bba_K1951004 digested E/P in pSB1C3 at 16°C overnight ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
 
* Ligation of Bba_K1951004 digested E/P in pSB1C3 at 16°C overnight ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
 
NB : we wanted to transform a cadA mutant from KEIO bank so we can't use this biobrick in pSB1K3.
 
NB : we wanted to transform a cadA mutant from KEIO bank so we can't use this biobrick in pSB1K3.
 
* Starters from intermediate biobirck Bba_K1951001 (DesB), Bba_K1951002(DesC), Bba_K1951003(DesD) with Bba_B0034 (rbs) and Bba_K1951000(DesA),Bba_K1951001(DesB), Bba_K1951002(DesC), Bba_K1951003(DesD) with Bba_B0034 (rbs) + BBa_I0500(Prom).
 
* Starters from intermediate biobirck Bba_K1951001 (DesB), Bba_K1951002(DesC), Bba_K1951003(DesD) with Bba_B0034 (rbs) and Bba_K1951000(DesA),Bba_K1951001(DesB), Bba_K1951002(DesC), Bba_K1951003(DesD) with Bba_B0034 (rbs) + BBa_I0500(Prom).
−
* Made competante cells Tg1, culture from an over night starter have grown up from Abs(600nm)=0.05 until Abs(600nm)=0.457 at 37°C ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.231_:_Preparation_of_competent_bacteria_cells Protocol#1]).
+
* Made competent cells Tg1, culture from an over night starter have grown up from Abs(600nm)=0.05 until Abs(600nm)=0.457 at 37°C ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.231_:_Preparation_of_competent_bacteria_cells Protocol#1]).
 
</div>
 
</div>
 
<div id="2016-09-01" class="evday">
 
<div id="2016-09-01" class="evday">
Line 757: Line 487:
 
* Ligation of Bba_K1951004 (S/P) and Bba_K1951001 + RBS (X/P) in pSB1K3(E/P) at 16°C overnight ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
 
* Ligation of Bba_K1951004 (S/P) and Bba_K1951001 + RBS (X/P) in pSB1K3(E/P) at 16°C overnight ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
 
* Ligation of Bba_K1951002 + RBS (E/S) and Bba_K1951003 + RBS (X/P) in pSB1K3(E/P) at 16°C overnight ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
 
* Ligation of Bba_K1951002 + RBS (E/S) and Bba_K1951003 + RBS (X/P) in pSB1K3(E/P) at 16°C overnight ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
−
* Made competante cell of fliC mutant and cadA mutant from the Keio bank and test on different antibiotic petri dishes (only kanamycin dish was a positive control) ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.231_:_Preparation_of_competent_bacteria_cells Protocol#1]).
+
* Made competent cells of fliC mutant and cadA mutant from the Keio bank and test on different antibiotic petri dishes (only kanamycin dish was a positive control) ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.231_:_Preparation_of_competent_bacteria_cells Protocol#1]).
 
* Retest of colony PCR on Bba_K1951001(DesB), Bba_K1951002(DesC), Bba_K1951003(DesD) with Bba_B0034 (rbs) + BBa_I0500(Prom) and nothing was good ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).
 
* Retest of colony PCR on Bba_K1951001(DesB), Bba_K1951002(DesC), Bba_K1951003(DesD) with Bba_B0034 (rbs) + BBa_I0500(Prom) and nothing was good ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).
−
* Transformation of the ligation Bba_K1951001(DesB), Bba_K1951002(DesC), Bba_K1951003(DesD) with Bba_B0034 (rbs) + BBa_I0500(Prom) from the ligation of the 08.26.2016 with the new competante from 08.30.2016 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
+
* Transformation of the ligation Bba_K1951001(DesB), Bba_K1951002(DesC), Bba_K1951003(DesD) with Bba_B0034 (rbs) + BBa_I0500(Prom) from the ligation of the 08.26.2016 with the new competent cells from 08.30.2016 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
−
* Transformation of Bba_K1951008 in fliC mutant Keio competante made the 30th, spread on Kana/Cm petri dishes ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
+
* Transformation of Bba_K1951008 in fliC mutant Keio competent made the 30th, spread on Kana/Cm petri dishes ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
 
* Made a calibration range for HPLC.
 
* Made a calibration range for HPLC.
 
</div>
 
</div>
Line 768: Line 498:
  
 
* Remade ligation of pSB1C3 (E/P) and Bba_K1951004 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
 
* Remade ligation of pSB1C3 (E/P) and Bba_K1951004 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
−
* Transformation of the previous ligation mix in competant Tg1 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
+
* Transformation of the previous ligation mix in competent Tg1 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
 
* Colony PCR on transformation of intermediate biobricks ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).:
 
* Colony PCR on transformation of intermediate biobricks ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).:
 
- Bba_K1951004 + RBS (X/P) Bba_K1951001 in pSB1K3(E/P) at 16°C overnight
 
- Bba_K1951004 + RBS (X/P) Bba_K1951001 in pSB1K3(E/P) at 16°C overnight
Line 787: Line 517:
 
=== Tuesday, September 6th ===
 
=== Tuesday, September 6th ===
 
[[File:T--Aix-Marseille--PCR-06-09.jpg|left|thumb|PCR colony verification. The holes 2 to 7 are the clone 12 of the plate and 11 to 16 are the clone 13 of the plate. The holes 9 and 17 are the RPF (control).]]
 
[[File:T--Aix-Marseille--PCR-06-09.jpg|left|thumb|PCR colony verification. The holes 2 to 7 are the clone 12 of the plate and 11 to 16 are the clone 13 of the plate. The holes 9 and 17 are the RPF (control).]]
−
* Transformation of cadA mutant competant strain by Bba_K1951004 in pSB1C3, spread on Kana/Cm petri dishes ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
+
* Transformation of cadA mutant competent strain by Bba_K1951004 in pSB1C3, spread on Kana/Cm petri dishes ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
 
* Colony PCR Bba_K1951004 in pSB1C3 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).
 
* Colony PCR Bba_K1951004 in pSB1C3 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).
 
* Starters from Bba_K1951004 in pSB1C3.
 
* Starters from Bba_K1951004 in pSB1C3.
 
* 1 step of the transduction using phage p1 from fliC mutant Keio bank. Conserved at 4°C with 20µL chlorophorme
 
* 1 step of the transduction using phage p1 from fliC mutant Keio bank. Conserved at 4°C with 20µL chlorophorme
 
* PCR with Q5 master mix using oligo of diriged mutagenesis for the insertion of Bbs1 site in the 214-215 and 238-239 sites of BBa_K1951008. Digest 3h with addition of 1µL DpNI in the PCR tube ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).
 
* PCR with Q5 master mix using oligo of diriged mutagenesis for the insertion of Bbs1 site in the 214-215 and 238-239 sites of BBa_K1951008. Digest 3h with addition of 1µL DpNI in the PCR tube ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).
−
* Transformation of the previous PCR in competant Tg1 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
+
* Transformation of the previous PCR in competent Tg1 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
 
* Starters of the PCR from 09.02.2016.
 
* Starters of the PCR from 09.02.2016.
 
* Remade PCR on Bba_K1951001(DesB), Bba_K1951002(DesC), Bba_K1951003(DesD) with Bba_B0034 (rbs) + BBa_I0500(Prom) but still no result ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).
 
* Remade PCR on Bba_K1951001(DesB), Bba_K1951002(DesC), Bba_K1951003(DesD) with Bba_B0034 (rbs) + BBa_I0500(Prom) but still no result ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_PCR_and_electrophoresis Protocol#3]).
Line 813: Line 543:
 
* Digestion of intermediate biobrick BBa_K1951004+ RBS BBa_K1951001 by EcoRI and SpeI and RBS BBa_K1951001+ RBS BBa_K1951002 by SpeI and PstI ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
 
* Digestion of intermediate biobrick BBa_K1951004+ RBS BBa_K1951001 by EcoRI and SpeI and RBS BBa_K1951001+ RBS BBa_K1951002 by SpeI and PstI ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
 
* Ligation of the 2 previous digestions in pSB1C3 to make the final BBa_K1951011 (E/P) ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
 
* Ligation of the 2 previous digestions in pSB1C3 to make the final BBa_K1951011 (E/P) ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.233_:_Cloning_protocol_for_IDT_sequences Protocol#5]).
−
* Transformation of the previous ligation in competant Tg1 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
+
* Transformation of the previous ligation in competent Tg1 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
 
</div>
 
</div>
 
<div id="2016-09-09" class="evday">
 
<div id="2016-09-09" class="evday">
Line 836: Line 566:
 
=== Sunday, September 11st ===
 
=== Sunday, September 11st ===
  
−
* Made competant of the fliC mutant W3110 strain from 5 differents colonies from 4 colonies which swam well ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.231_:_Preparation_of_competent_bacteria_cells Protocol#1]).     
+
* Made competent of the fliC mutant W3110 strain from 5 differents colonies from 4 colonies which swam well ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.231_:_Preparation_of_competent_bacteria_cells Protocol#1]).     
−
* Transformation of the previous competant with BBa_K1951008 and BBa_K1951009 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
+
* Transformation of the previous competent with BBa_K1951008 and BBa_K1951009 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
 
* Transformation of cadA mutant Keio strain with BBa_K1951011 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
 
* Transformation of cadA mutant Keio strain with BBa_K1951011 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
 
</div>
 
</div>
Line 844: Line 574:
 
=== Monday, September 12rd ===
 
=== Monday, September 12rd ===
 
    
 
    
−
* Transformation of the fliC mutant W3110 competant with BBa_K1951008 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
+
* Transformation of the fliC mutant W3110 competent with BBa_K1951008 ([https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols#Protocol_.232_:_Transformations Protocol#2]).
 
</div>
 
</div>
 
<div id="2016-09-13" class="evday">
 
<div id="2016-09-13" class="evday">

Latest revision as of 20:04, 19 October 2016