Difference between revisions of "Team:Stuttgart/Notebook"

Line 1: Line 1:
 +
{{Template:Home}}
 
<html>
 
<html>
−
 
−
<!--- THIS IS WHERE THE HTML BEGINS --->
 
−
 
−
<head>
 
−
    <link href="https://netdna.bootstrapcdn.com/bootstrap/3.0.0/css/bootstrap.min.css" rel="stylesheet"
 
−
          id="bootstrap-css">
 
−
    <style>
 
−
 
−
 
−
        /***************************************************** DEFAULT WIKI SETTINGS  ****************************************************/
 
−
 
−
        /* Clear the default wiki settings */
 
−
 
−
        #home_logo, #sideMenu {
 
−
            display: none;
 
−
        }
 
−
 
−
        #sideMenu, #top_title, .patrollink {
 
−
            display: none;
 
−
        }
 
−
 
−
        #content {
 
−
            width: 100%;
 
−
            padding: 0px;
 
−
            margin-top: -7px;
 
−
            margin-left: 0px;
 
−
        }
 
−
 
−
        body {
 
−
            background: white;
 
−
        }
 
−
 
−
        #bodyContent h1, #bodyContent h2, #bodyContent h3, #bodyContent h4, #bodyContent h5 {
 
−
            margin-bottom: 0;
 
−
            color: #F8CE63;
 
−
        }
 
−
 
−
        table {;
 
−
        }
 
−
 
−
        /**************************************************************** MENU ***************************************************************/
 
−
        /* Wrapper for the menu */
 
−
        .igem_2017_menu_wrapper {
 
−
            width: 15%;
 
−
            height: 100vh;
 
−
            position: fixed;
 
−
            right: 0;
 
−
            padding: 0;
 
−
            float: right;
 
−
            border-left: 1px solid #F8CE63;
 
−
            background-color: white;
 
−
            text-align: left;
 
−
            font-family: Tahoma, Geneva, sans-serif;
 
−
            overflow-y: auto;
 
−
            overflow-x: hidden;
 
−
        }
 
−
 
−
        /* this hides the scrollbar to keep view consistency */
 
−
        .igem_2017_menu_wrappe::-webkit-scrollbar {
 
−
            display: none;
 
−
        }
 
−
 
−
        /* styling for links in the menu, removes the line under text */
 
−
        .igem_2017_menu_wrapper a {
 
−
            text-decoration: none;
 
−
        }
 
−
 
−
        /* styling for the images in the menu */
 
−
        .igem_2017_menu_wrapper img {
 
−
            width: 100%;
 
−
        }
 
−
 
−
        /* styling for the menu buttons */
 
−
        .igem_2017_menu_wrapper .menu_button {
 
−
            width: 100%;
 
−
            padding: 10px 0px 10px 15px;
 
−
            float: left;
 
−
            border-bottom: 1px solid #F8CE63;
 
−
            font-size: 16px;
 
−
            font-weight: bold;
 
−
            color: #F8CE63;
 
−
            cursor: pointer;
 
−
        }
 
−
 
−
        .igem_2017_menu_wrapper .menu_bottom_padding {
 
−
            width: 100%;
 
−
            height: 30px;
 
−
            float: left;
 
−
        }
 
−
 
−
        .menu_button.direct_to_page {
 
−
            padding-left: 36px;
 
−
        }
 
−
 
−
        .igem_2017_menu_wrapper .menu_button .expand_collapse_icon {
 
−
            width: 10%;
 
−
            float: left;
 
−
        }
 
−
 
−
        .igem_2017_menu_wrapper .menu_button .expand_collapse_icon::before {
 
−
            content: "+";
 
−
        }
 
−
 
−
        /* styling for the menu buttons on hover */
 
−
        .igem_2017_menu_wrapper .menu_button:hover, .igem_2017_menu_wrapper .submenu_button:hover, .submenu_button.current_page:hover {
 
−
            background-color: #F8CE63;
 
−
            text-decoration: none;
 
−
            color: #190707;
 
−
        }
 
−
 
−
        /* styling for the menu button when it is the current page */
 
−
        .current_page {
 
−
            background-color: white !important;
 
−
            color: #F8CE63 !important;
 
−
        }
 
−
 
−
        /* styling for the submenu buttons */
 
−
        .igem_2017_menu_wrapper .submenu_button {
 
−
            width: 100%;
 
−
            padding: 10px 0px 10px 34px;
 
−
            float: left;
 
−
            background-color: white;
 
−
            border-bottom: 1px solid #F8CE63;
 
−
            font-size: 15px;
 
−
            color: #F8CE63;
 
−
            cursor: pointer;
 
−
        }
 
−
 
−
        /* wrapper for the submenu items, they are hidden by default*/
 
−
        .igem_2017_menu_wrapper .submenu_wrapper {
 
−
            display: none;
 
−
        }
 
−
 
−
        /* when the page size is bigger than 800px, this show/hide control is hidden by default */
 
−
        .igem_2017_menu_wrapper #display_menu_control {
 
−
            display: none;
 
−
            text-align: center;
 
−
        }
 
−
 
−
        /***************************************************** CONTENT OF THE PAGE ****************************************************/
 
−
 
−
        /* Wrapper for the content */
 
−
        .igem_2017_content_wrapper {
 
−
            width: 83%;
 
−
            margin: 0;
 
−
            display: block;
 
−
            float: left;
 
−
            background-color: white;
 
−
            font-family: Tahoma, Geneva, sans-serif;
 
−
        }
 
−
 
−
        /********************************* HTML STYLING  *********************************/
 
−
 
−
        /* styling for the titles h1 h2 */
 
−
        .igem_2017_content_wrapper h1, .igem_2017_content_wrapper h2 {
 
−
            color: #F8CE63;
 
−
            padding: 15px 0 25px 0;
 
−
            border-bottom: 0;
 
−
        }
 
−
 
−
        /* styling for the titles  h3 h4 h5 h6*/
 
−
        .igem_2017_content_wrapper h3, .igem_2017_content_wrapper h4, .igem_2017_content_wrapper h5, .igem_2017_content_wrapper h6 {
 
−
            padding: 5px 15px;
 
−
            border-bottom: 0;
 
−
        }
 
−
 
−
        /* font and text */
 
−
        .container p,
 
−
        #HQ_page p {
 
−
            line-height: 24px;
 
−
            font-size: 16px;
 
−
            color: #333;
 
−
        }
 
−
 
−
        .igem_2017_content_wrapper img {
 
−
            padding: 15px 0;
 
−
        }
 
−
 
−
        /* Links */
 
−
        .igem_2017_content_wrapper a {
 
−
            font-weight: bold;
 
−
            text-decoration: underline;
 
−
            text-decoration-color: #190707;
 
−
            color: #3399ff;
 
−
            -webkit-transition: all 0.4s ease;
 
−
            -moz-transition: all 0.4s ease;
 
−
            -ms-transition: all 0.4s ease;
 
−
            -o-transition: all 0.4s ease;
 
−
            transition: all 0.4s ease;
 
−
        }
 
−
 
−
        /* hover for the links */
 
−
        .igem_2017_content_wrapper a:hover {
 
−
            text-decoration: none;
 
−
            color: #000000;
 
−
        }
 
−
 
−
        /* non numbered lists */
 
−
        .igem_2017_content_wrapper ul {
 
−
            padding: 0px 20px;
 
−
            font-size: 15px;
 
−
            font-family: Tahoma, Geneva, sans-serif;
 
−
        }
 
−
 
−
        /* numbered lists */
 
−
        .igem_2017_content_wrapper ol {
 
−
            padding: 0px;
 
−
            font-size: 15px;
 
−
            font-family: Tahoma, Geneva, sans-serif;
 
−
        }
 
−
 
−
        /* Table */
 
−
        .igem_2017_content_wrapper table {
 
−
            width: 97%;
 
−
            margin: 15px 10px;
 
−
            border: 2px solid #190707;
 
−
            border-collapse: collapse;
 
−
        }
 
−
 
−
        /* table cells */
 
−
        .igem_2017_content_wrapper td {
 
−
            padding: 10px;
 
−
            vertical-align: text-top;
 
−
            border: 1px solid #190707;
 
−
            border-collapse: collapse;
 
−
        }
 
−
 
−
        /* table headers */
 
−
        .igem_2017_content_wrapper th {
 
−
            padding: 10px;
 
−
            vertical-align: text-top;
 
−
            border: 1px solid #190707;
 
−
            border-collapse: collapse;
 
−
 
−
        }
 
−
 
−
        /**********************************LAYOUT CLASSES **********************************/
 
−
 
−
        /* general class for column divs */
 
−
        .igem_2017_content_wrapper .column {
 
−
            padding: 10px 0px;
 
−
            float: left;
 
−
        }
 
−
 
−
        /* class for a full width column */
 
−
        .column .full_size {
 
−
            width: 100%;
 
−
        }
 
−
 
−
        /* styling for images in a full width column*/
 
−
        .column.full_size img {
 
−
            width: 97%;
 
−
            padding: 10px 15px;
 
−
        }
 
−
 
−
        /* class for a half width column */
 
−
        .column.half_size {
 
−
            width: 50%;
 
−
        }
 
−
 
−
        /* styling for images in a half width column*/
 
−
        .column.half_size img {
 
−
            width: 94.5%;
 
−
            padding: 10px 15px;
 
−
        }
 
−
 
−
        /********************************* SUPPORT CLASSES ********************************/
 
−
 
−
        /* class that clears content below*/
 
−
        .igem_2017_content_wrapper .clear {
 
−
            clear: both;
 
−
        }
 
−
 
−
        /* adds extra spacing when clearing content */
 
−
        .igem_2017_content_wrapper .clear.extra_space {
 
−
            height: 30px;
 
−
        }
 
−
 
−
        /* highlight class, makes content slightly smaller */
 
−
        .igem_2017_content_wrapper .highlight {
 
−
            margin: 0px 15px;
 
−
            padding: 15px 0px;
 
−
        }
 
−
 
−
        /* highlight class, adds a gray background */
 
−
        .igem_2017_content_wrapper .highlight.gray {
 
−
            background-color: #190707;
 
−
        }
 
−
 
−
        /* highlight with decoration blue line on top */
 
−
        .igem_2017_content_wrapper .highlight.blue_top {
 
−
            border-top: 4px solid #190707;
 
−
        }
 
−
 
−
        /* highlight with a full blue border decoration */
 
−
        .igem_2017_content_wrapper .highlight.blue_border {
 
−
            border: 4px solid #190707;
 
−
        }
 
−
 
−
        /* button class */
 
−
        .igem_2017_content_wrapper .button {
 
−
            max-width: 35%;
 
−
            margin: 30px auto;
 
−
            padding: 12px 10px;
 
−
            background-color: #190707;
 
−
            text-align: center;
 
−
            color: #F8CE63;
 
−
            -webkit-transition: all 0.4s ease;
 
−
            -moz-transition: all 0.4s ease;
 
−
            -ms-transition: all 0.4s ease;
 
−
            -o-transition: all 0.4s ease;
 
−
            transition: all 0.4s ease;
 
−
        }
 
−
 
−
        /* styling for button on hover */
 
−
        .igem_2017_content_wrapper .button:hover {
 
−
            background-color: #190707;
 
−
            color: #F8CE63;
 
−
        }
 
−
 
−
        /***************************************************** RESPONSIVE STYLING ****************************************************/
 
−
 
−
        *, *:before, *:after {
 
−
            -webkit-box-sizing: border-box;
 
−
            -moz-box-sizing: border-box;
 
−
            box-sizing: border-box;
 
−
        }
 
−
 
−
        body {
 
−
            overflow-x: hidden;
 
−
            margin-top: 80px;
 
−
            letter-spacing: 0.02em;
 
−
        }
 
−
 
−
        /* IF THE SCREEN IS LESS THAN 1200PX */
 
−
        @media only screen and (max-width: 1024px) {
 
−
 
−
            #content {
 
−
                width: 100%;
 
−
            }
 
−
 
−
            .igem_2017_content_wrapper {
 
−
                width: 100%;
 
−
            }
 
−
 
−
            .igem_2017_menu_wrapper {
 
−
                width: 100%;
 
−
                height: 15%;
 
−
                position: relative;
 
−
                left: 0;
 
−
            }
 
−
 
−
            .highlight {
 
−
                padding: 10px 0px;
 
−
            }
 
−
 
−
            .igem_2017_menu_wrapper #display_menu_control {
 
−
                display: none;
 
−
            }
 
−
 
−
            #menu_content {
 
−
                display: block;
 
−
            }
 
−
 
−
            .menu_button.direct_to_page {
 
−
                padding-left: 17px;
 
−
            }
 
−
        }
 
−
 
−
        /* IF THE SCREEN IS LESS THAN 800PX */
 
−
        @media only screen and (max-width: 800px) {
 
−
 
−
            .igem_2017_content_wrapper {
 
−
                width: 100%;
 
−
                margin-left: 0;
 
−
            }
 
−
 
−
            .column.half_size {
 
−
                width: 100%;
 
−
            }
 
−
 
−
            .column.full_size img, .column.half_size img {
 
−
                width: 100%;
 
−
                padding: 10px 0;
 
−
            }
 
−
 
−
            .highlight {
 
−
                padding: 15px 5px;
 
−
            }
 
−
 
−
            .igem_2017_menu_wrapper #display_menu_control {
 
−
                display: block;
 
−
            }
 
−
 
−
            #menu_content {
 
−
                display: none;
 
−
            }
 
−
 
−
            .igem_2017_menu_wrapper .menu_button .expand_collapse_icon {
 
−
                width: 5%;
 
−
            }
 
−
 
−
            .menu_bottom_padding {
 
−
                display: none;
 
−
            }
 
−
 
−
            .menu_button.direct_to_page {
 
−
                padding-left: 36px;
 
−
            }
 
−
        }
 
−
 
−
        /* special class that the system uses to make sure the team wants a page to be evaluated */
 
−
        .judges-will-not-evaluate {
 
−
            width: 96.6%;
 
−
            margin: 5px 15px;
 
−
            display: block;
 
−
            border: 4px solid #3399ff;
 
−
            font-weight: bold;
 
−
        }
 
−
 
−
        /* CUSTOM BS NAVBAR STYLES */
 
−
        .navbar {
 
−
            border: 0;
 
−
        }
 
−
 
−
        .navbar-default {
 
−
            margin-top: 15px;
 
−
            background-color: #333;
 
−
        }
 
−
 
−
        .navbar-brand {
 
−
            padding: 0;
 
−
            padding-left: 25px;
 
−
            padding-right: 25px;
 
−
        }
 
−
 
−
        .navbar-brand img {
 
−
            height: 49px;
 
−
        }
 
−
 
−
        .navbar-default .navbar-nav > li > a {
 
−
            color: white;
 
−
        }
 
−
 
−
        .navbar-default .navbar-nav > li > a:focus,
 
−
        .navbar-default .navbar-nav > li > a:hover,
 
−
        .navbar-default .navbar-nav > .dropdown > a .caret:hover {
 
−
            color: #F8CE63;
 
−
        }
 
−
 
−
        .navbar-default .navbar-nav > .open > a,
 
−
        .navbar-default .navbar-nav > .open > a:hover,
 
−
        .navbar-default .navbar-nav > .open > a:focus {
 
−
            color: #F8CE63;
 
−
            background-color: #444;
 
−
        }
 
−
 
−
        .navbar-default .navbar-nav > .dropdown > a .caret {
 
−
            border-top-color: #eee;
 
−
            border-bottom-color: #eee;
 
−
        }
 
−
 
−
        .dropdown-menu {
 
−
            border: 0;
 
−
        }
 
−
        .dropdown-menu > li > a {
 
−
            color: white;
 
−
        }
 
−
 
−
        .dropdown-menu > li > a:hover,
 
−
        .dropdown-menu > li > a:focus {
 
−
            color: #111;
 
−
            text-decoration: none;
 
−
            background-color: #F8CE63;
 
−
        }
 
−
 
−
        .navbar-default .navbar-nav > .dropdown > a:hover .caret,
 
−
        .navbar-default .navbar-nav > .dropdown > a:focus .caret {
 
−
            border-top-color: white;
 
−
            border-bottom-color: white;
 
−
        }
 
−
 
−
        @media (max-width: 767px) {
 
−
            .navbar-default .navbar-nav .open .dropdown-menu>li>a {
 
−
                color: #ddd;
 
−
            }
 
−
            .navbar-default .navbar-nav .open .dropdown-menu>li>a:focus,
 
−
            .navbar-default .navbar-nav .open .dropdown-menu>li>a:hover {
 
−
                color: #F8CE63;
 
−
            }
 
−
        }
 
−
 
−
        .navbar-nav>li>.dropdown-menu {
 
−
            background-color: #444;
 
−
        }
 
−
 
−
        .section {
 
−
            margin: 25px 0;
 
−
        }
 
−
 
−
    </style>
 
−
    <script src="https://code.jquery.com/jquery-1.10.2.min.js"></script>
 
−
    <script src="https://netdna.bootstrapcdn.com/bootstrap/3.0.0/js/bootstrap.min.js"></script>
 
−
 
−
    <script>
 
−
        $(document).ready(function(){
 
−
            $("#mw-content-text").removeClass("mw-content-ltr");
 
−
        });
 
−
    </script>
 
−
 
−
 
−
    <!-- This tells the browser that your page is responsive -->
 
−
    <meta name="viewport" content="width=device-width, initial-scale=1">
 
−
 
−
</head>
 
−
<body>
 
−
 
−
<!--MENU-->
 
−
<nav class="navbar navbar-default navbar-fixed-top">
 
−
    <div class="container-fluid">
 
−
        <!-- Brand and toggle get grouped for better mobile display -->
 
−
        <div class="navbar-header">
 
−
            <button type="button" class="navbar-toggle collapsed" data-toggle="collapse"
 
−
                    data-target="#bs-example-navbar-collapse-1" aria-expanded="false">
 
−
                <span class="sr-only">Toggle navigation</span>
 
−
                <span class="icon-bar"></span>
 
−
                <span class="icon-bar"></span>
 
−
                <span class="icon-bar"></span>
 
−
            </button>
 
−
            <a class="navbar-brand" href="https://2017.igem.org/Team:Stuttgart">
 
−
                <img alt="Team:Stuttgart"
 
−
                    src="https://static.igem.org/mediawiki/2017/1/18/UNISTUTTGARTLOGO.png">
 
−
            </a>
 
−
        </div>
 
−
 
−
        <!-- Collect the nav links, forms, and other content for toggling -->
 
−
        <div class="collapse navbar-collapse" id="bs-example-navbar-collapse-1">
 
−
            <ul class="nav navbar-nav">
 
−
                <li>
 
−
                    <a href="https://2017.igem.org/Team:Stuttgart">Home</a>
 
−
                </li>
 
−
                <li class="dropdown">
 
−
                    <a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-haspopup="true"
 
−
                      aria-expanded="false">Team <span class="caret"></span></a>
 
−
                    <ul class="dropdown-menu">
 
−
                        <li><a href="https://2017.igem.org/Team:Stuttgart/Team">Team</a></li>
 
−
                        <li><a href="https://2017.igem.org/Team:Stuttgart/Attributions">Attributions</a></li>
 
−
                        <li><a href="https://2017.igem.org/Team:Stuttgart/Sponsoring">Sponoring</a></li>
 
−
                        <!--<li role="separator" class="divider"></li>-->
 
−
                    </ul>
 
−
                </li>
 
−
                <li class="dropdown">
 
−
                    <a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-haspopup="true"
 
−
                      aria-expanded="false">Project <span class="caret"></span></a>
 
−
                    <ul class="dropdown-menu">
 
−
                        <!--<li class="navbar-text navbar-left">Dry Lab</li>-->
 
−
                        <li><a href="https://2017.igem.org/Team:Stuttgart/Description">Description</a></li>
 
−
                        <li><a href="https://2017.igem.org/Team:Stuttgart/Background-Information">Background
 
−
                            Information</a></li>
 
−
                        <li><a href="https://2017.igem.org/Team:Stuttgart/Protocols">Protocols & Experiments</a></li>
 
−
                        <li><a href="https://2017.igem.org/Team:Stuttgart/Notebook">Notebook</a></li>
 
−
                        <li><a href="https://2017.igem.org/Team:Stuttgart/InterLab">Interlab Study</a></li>
 
−
                        <li><a href="https://2017.igem.org/Team:Stuttgart/Model">Modelling</a></li>
 
−
                        <li><a href="https://2017.igem.org/Team:Stuttgart/Results">Results</a></li>
 
−
                        <li><a href="https://2017.igem.org/Team:Stuttgart/Parts">Parts</a></li>
 
−
                        <!--<li role="separator" class="divider"></li>-->
 
−
                    </ul>
 
−
                </li>
 
−
                <li>
 
−
                    <a href="https://2017.igem.org/Team:Stuttgart/Safety">Safety</a>
 
−
                </li>
 
−
                <li class="dropdown">
 
−
                    <a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-haspopup="true"
 
−
                      aria-expanded="false">Human Practices <span class="caret"></span></a>
 
−
                    <ul class="dropdown-menu">
 
−
                        <li><a href="https://2017.igem.org/Team:Stuttgart/Engagement">Public Engagement</a></li>
 
−
                        <li><a href="https://2017.igem.org/Team:Stuttgart/HP/Gold_Integrated">Integrated and Gold</a>
 
−
                        </li>
 
−
                        <li><a href="https://2017.igem.org/Team:Stuttgart/HP/Silver">Silver HP</a></li>
 
−
                        <li><a href="https://2017.igem.org/Team:Stuttgart/Collaborations">Collaborations</a></li>
 
−
                    </ul>
 
−
                </li>
 
−
                <li class="dropdown">
 
−
                    <a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-haspopup="true"
 
−
                      aria-expanded="false">Awards <span class="caret"></span></a>
 
−
                    <ul class="dropdown-menu">
 
−
                        <li><a href="https://2017.igem.org/Team:Stuttgart/Applied_Design">Applied Design</a></li>
 
−
                        <li><a href="https://2017.igem.org/Team:Stuttgart/Measurement">Measurement</a></li>
 
−
                    </ul>
 
−
                </li>
 
−
                <li>
 
−
                    <a href="https://igem.org/2017_Judging_Form?team=Stuttgart">Judging Form</a>
 
−
                </li>
 
−
            </ul>
 
−
        </div><!-- /.navbar-collapse -->
 
−
    </div><!-- /.container-fluid -->
 
−
</nav>
 
−
 
−
 
−
 
−
 
  
 
<!-- start of content -->
 
<!-- start of content -->
Line 796: Line 196:
 
</div>
 
</div>
  
−
</body>
 
 
</html>
 
</html>

Revision as of 19:20, 29 October 2017

Notebook

Esterases and Lipases


03.08.2017

  • Transformation of BBa_K1149002 and BBa_K1149003

04.08.17

  • Single colonies (BBa_K1149002 and BBa_K1149003) are plated on agar plates and incubated at 37 °C over night

09.08.17

  • Growing of a single colony (BBa_K1149002 and BBa_K1149003) in 5 mL LB media + chloramphenicol (35 µg/mL) at 37 °C

10.08.17

  • Preparation of miniprep (Jena Biosciences Kit) and glycerol stock (storage: -80 °C) (BBa_K1149002 and BBa_K1149003)
  • SOC media preparation
  • Transformation pet19-LipB

21.08.2017

  • Preparation: preliminary test of esterase assay with EstCS2: Bba_K1149002 - preparation of o/n cultures (37°C)

22.08.2017

  • Preparation: preliminary test of esterase assay with EstCS2: Bba_K1149002 - glycerol stocks and induction with arabinose

23.08.2017

  • Preliminary test of esterase assay with EstCS2: Bba_K1149002 (link results)

28.08.2017

  • Preparation of electro-competent cells

31.08.2017

  • Transformation/electroporation with iGEM competent cells test kit DNA (o/n incubation, 37°C - transformation failed)

01.09.2017

  • Transformation/electroporation with pUC19 plasmid (o/n incubation, 37°C - transformation failed)

06.09.2017-08.09.17

  • Enzyme activity assay (cell pellet and supernatant) with BBa_K1149002 and pet19-LipB

12.09.2017

  • Preparation of chemo-competent cells

13.09.2017

  • Transformation with iGEM competent cells test kit DNA and pUC19 plasmid (o/n incubation, 37°C) - transformation successful


14.09.2017

  • Calculation - efficiency of the chemo-competent cells/pUC19: efficiency E =(56cfu/0.001ng)*1000ng/(µL)=5.6*((10)^7 )cfu/µL

18.09.2017<

  • Preparation InterLab study - transformation of 8 parts into DH5alph E. coli cells

19.09.17 – 21.09.17

  • InterLab study LUDOX measurement
  • Enzyme activity assay with BBa_K1149002 of the supernatant - measurement in biological triplicates
  • InterLab study Fluorescein measurement

22.09.2017

  • InterLab study sample measurement (link results)

27.09.17 - 29.09.17

  • Enzyme activity assay with pet19-LipB of the supernatant - measurement in biological triplicates

06.10.2017

  • Collaboration iGEM team Heidelberg: preparation mutagenesis plasmid activity assay - preparation of media, antibiotic-stocks and sugar-stocks + agar plates with different antibiotics

10.10.2017

  • Collaboration iGEM team Heidelberg: preparation of different cultures + induction of the cells with arabinose (o/n incubation, 37°C)

11.10.2017

  • Collaboration iGEM team Heidelberg: spread the o/n cultures on prepared agar plates (o/n incubation, 37°C)

12.10.2017

  • Gibson Assembly of LipB in psB1C3 backbone

19.10.2017

  • PCR - amplification of LipB - cloning of LipB in the iGEM standard plasmid (BBa_K1149002 is used)
  • PCR - amplification of BBa_K1149002 (without EstCS2)

20.10.2017

  • SDS page, Gibson Assembly, Transformation (Zymos) - cloning of LipB in the iGEM standard plasmid (BBa_K1149002 is used)

Keratinases


26.07.2017

  • Transformation of kerUS (BBa_K1498000)
  • Preparation of antibiotic stock solutions: chloramphenicol (35 mg/mL) and ampicillin (100 mg/mL)

27.07.2017

  • Growing of a single colony (kerUS (BBa_K1498000)) in 5 mL LB media chloramphenicol (35 µg/mL) at 37 °C
  • Single colonies are plated on agar plates and incubated at 37 °C over night

28.07.2017

  • Preparation of: miniprep (Jena Biosciences Kit)and glycerol stock (kerUS (BBa_K1498000))

30.08.17

  • Transformation of psB1K3-KerP

12.10.2017 <

  • Gibson Assembly KerA-ompA, ompA-KerA, KerUS-ompA and ompA-KerUS in psB1C3 backbone

16.10.2017

  • Preparation semi-quantitative keratinase-assay: spread kerUS, KerA and KerP on chloramphenicol/kanamycin agar plates (o/n incubation, 37°C)
  • Preparation skim-milk-plate assay: preparation of skim-milk-agar-plates with chloramphenicol/kanamycin

18.10.2017

  • Preparation semi-quantitative keratinase-assay: preparation of o/n cultures (37°C)
  • Preparation skim-milk-plate assay: preparation of o/n cultures (37°C)

19.10.2017

  • Semi-quantitative keratinase-assay: add 0.005 g of dried hair (65 °C, 1h) to o/n cultures (KerUS and KerA are induced with 1mM IPTG before) - 5 days incubation, 37°C (link results)
  • Skim-milk-plate assay kerP - spread on prepared skim-milk agar plates (supernatant and cells) - 4 days incubation, room temperature (link results)

20.10.2017

  • Skim-milk-plate assay kerUS and KerA - spread on prepared skim-milk agar plates (supernatant and cells) - 4 days incubation, room temperature (link results)
  • Sequencing of keratinase plasmids: pSB1C3-KerA-ompA, pSB1C3-KerUS-ompA, pSB1C3-KerA_Kanada and pSB1C3-KerUS-Kanada< /li>

Rose and Limonene Fragrance


28.08.2017

  • Preparation of LB-Agar plates with kanamycin (50 µg/mL)
  • Sequencing of rose fragrance plasmids from Guo et al. (pET28a-KDC-YjgB-ARO8 and pET28a-ATF1)

13.09.2017

  • Overlap-PCR of 1.pBAD (BBa_K206000), 2.KDC-YjgB-ARO8, 3.ATF1 and 4.pBAD-KDC-YjgB-ARO8
  • PCR-Purification and agarose-gel-electrophoresis of PCR products
  • PCR 4 (pBAD-KDC-YjgB-ARO8) not successful

19.09.2017

  • Preparation of LB-Agar plates with kanamycin (50 µg/mL) and chloramphenicol (35 µg/mL)
  • Repeat of PCR 4 (pBAD-KDC-YjgB-ARO8)
  • PCR-Purification and agarose-gel-electrophoresis of PCR product 4

  • PCR 4 (pBAD-KDC-YjgB-ARO8) not successful
  • Transformation of Limonene-plasmid and over-night incubation on agar-plates with kanamycin (50 µg/mL) at 37°C

21.09.17

  • Gibson Assembly of rose PCR-overlap products pBAD, KDC-YjgB-ARO8 and ATF1 in psB1K3 backbone
  • Transformation of assembled rose-plasmid and over-night incubation on agar-plates with kanamycin (50 µg/mL) at 37°C

22.09.2017

  • Rose-plasmid Transformation successful
  • Single colonies are plated on agar plates and incubated at 37 °C over night

25.09.2017

  • Verification of transformed rose-plasmid by colony-PCR
  • Agarose-gel-electrophoresis of colony-PCR product - colony-PCR not successful

26.09.2017

  • Repeat of colony-PCR with transformed rose-plasmid
  • Agarose-gel-electrophoresis of colony-PCR product - colony-PCR not successful

27.09.2017

  • Repeat of colony-PCR with transformed rose-plasmid
  • Agarose-gel-electrophoresis of colony-PCR product - colony-PCR not successful

28.09.2017

  • Mini-prep of transformed rose-plasmid
  • Restriction assay of isolated rose-plasmid, cut with SpeI
  • Verification of restriction product by agarose-gel-electrophoresis - not successful

29.09.2017

  • Repeat Gibson Assembly of rose PCR-overlap products pBAD, KDC-YjgB-ARO8 and ATF1
  • Transformation of assembled rose-plasmid and over-night incubation on agar-plates with kanamycin (50 µg/mL) at 37°C - transformation not successful

05.10.2017

  • Repeat Restriction assay of isolated rose-plasmid, cut with EcoRI - not successful
  • Verification of restriction product by agarose-gel-electrophoresis - not successful

12.10.2017

  • Gibson Assembly of limonene PCR-products () in psB1C3 backbone
  • Transformation of assembled rose-plasmid and over-night incubation on agar-plates with kanamycin (50 µg/mL) at 37°C - transformation not successful

18.10.2017

  • Repeat of colony-PCR with transformed rose-plasmid, temperature range from 58-70 °C
  • Agarose-gel-electrophoresis of colony-PCR product - colony-PCR not successful
  • M9 media preparation

20.10.2017

  • Repeat overlap-PCR of pBad and KDC-YjgB-ARO8
  • Verification of PCR products by agarose-gel-electrophoresis

23.10.2017

  • Sequencing of transformed rose fragrance plasmid (pSB1K3-pBad-KDC-YjgB-ARO8-ATF1)
  • Sequencing of transformed limonene fragrance plasmid (pSB1C3-pBad)

25.10.2017

  • Repeat Gibson Assembly of rose PCR-overlap products pBAD, KDC-YjgB-ARO8 and ATF1, plasmid backbones pSB1K3 and pSB1C3
  • Transformation of assembled rose-plasmid and over-night incubation on agar-plates with kanamycin (50 µg/mL) and chloramphenicol (35 µg/mL) at 37°C - transformation not successful

26.10.2017

  • Repeat transfomation of rose-plasmid in competent NEB-cells and and over-night incubation on agar-plates with kanamycin (50 µg/mL) and chloramphenicol (35 µg/mL) at 37°C - transformation successful

27.10.2017

  • Verification of transformed rose-plasmid by colony-PCR
  • Agarose-gel-electrophoresis of colony-PCR product - colony-PCR not successful